Bethyl Laboratories

Olink AB and Bethyl sign distribution agreement for the US market

2011-11-21T05:38:00.000-08:00

UPPSALA, SWEDEN, November 2011 - Olink Bioscience, a Sweden based biotechnology company focused on innovative protein detection technologies, has entered into a distribution agreement with Bethyl Laboratories, Inc. Together with Bethyl Olink Bioscience will continue to expand the distribution of the Duolink® product line in the US market.

“We are excited to enter into an agreement with Bethyl to distribute our Duolink product line in the USA. With an excellent reputation as a provider of first class products we believe Bethyl will help us expand the market for our products in the region”, comments Simon Fredriksson, President and CEO, Olink AB.

“Bethyl is an established provider of premium quality antibody products and services for clinical and research applications and through the distribution agreement with Olink we now have the opportunity to offer a complete solution of antibodies together with a highly specific protein detection technology” says John Carwile, CEO, Bethyl Laboratories, Inc.

About Bethyl
Since 1972, Bethyl has provided quality antibody products and services for both clinical and research applications. We remain committed to manufacturing new and novel antibodies to facilitate and accelerate your discovery based research in cell biology. To ensure the quality of our antibodies, all antibodies are produced and affinity purified by antigen specific affinity chromatography at our sole facility in Montgomery, TX. Through our collaborations and scholarly searches we continue to identify new and emerging proteins of interests. We are aggressively producing antibodies to these new proteins as well as those classic proteins for which there is a dearth of quality antibodies. We test our antibodies in a wide range of applications including ELISA, western blot, immunoprecipitation, immunohistochemistry, immunocytochemistry and proximity ligation assay.

Bethyl Laboratories is a Registered Research Facility with the USDA under the Animal Welfare Act and complies daily with all requirements of the Act. Bethyl Laboratories, Inc. operates under Good Manufacturing Practices.

About Olink AB
Olink Bioscience, a company founded by scientists at Uppsala University, is commercializing cutting-edge technologies for analysis of proteins and nucleic acids revealing new insights into basic science, drug development, and diagnostics. Our products are available worldwide through a network of distributors and through our web shop. The Duolink® product line enables users to visualize and quantify individual proteins, their interactions and modifications, in unmodified cells and tissues. Proseek® is an open assay development reagent kit for analyzing proteins in only 1 µl of serum or plasma sample with high sensitivity.

Olink Bioscience molecular technologies are also commercialized through partnerships with industry leading organizations such as Affymetrix, Life Technologies, and through Olink spin-out companies Halo Genomics and Qlinea. Olink Bioscience was founded in 2004 and is headquartered in Uppsala, Sweden

For more information, please contact:

John M. Carwile, MD

Chief Executive Officer

Bethyl Laboratories, Inc.

(800) 338-9579 (main)

jcarwile@bethyl.com

Focusing on Homologous Recombination

2011-09-06T10:39:00.000-07:00

Double stranded breaks (DSBs) are a normal occurrence during DNA replication. The cellular response to DSBs is complex and highly orchestrated, and a cell’s inability to properly address DSBs can lead to genetic alterations such as loss of heterozygosity, mutations, deletions, genomic rearrangements, and chromosome loss. There are two major pathways for repairing DSBs: non-homologous end joining (NHEJ) and homologous recombination (HR). Both pathways are critical to the maintenance of genomic stability and involve a variety of proteins that play particular functional roles in the response to DNA damage. In HR, the functional groups include the DNA damage sensors, cell cycle checkpoint enforcers, and the catalyzers of HR repair itself. Additionally, the HR repair pathway requires proteins that function as adaptors and transducers, effector kinases, regulatory proteins, and structural components (1). There a re several diseases associated with genetic defects in proteins involved in HR. These genetic diseases are characterized by a predisposition to cancer indicating the importance of intact HR pathways to circumvent cancer (2). Although deficiencies in HR lead to cancer development, these deficiencies also allow for the selective treatment of cancer cells. Anti-cancer drugs mainly kill cancer cells by inducing toxic DSBs and defects in HR can increase the sensitivity of cancer cells to these drugs (3). Therefore, a better understanding of the factors and pathways involved in HR will further advancements in the development of therapies for cancer.

Selected Reviews

1. Lisby, M. & Rothstein, R. (2009). Choreography of recombination proteins during the DNA damage response. DNA Repair (Amst), 8, 1068-1076.

2. Thompson, L. H. & Schild, D. (2002). Recombinational DNA repair and human disease. Mutat.Res., 509, 49-78.

3. Helleday, T. (2010). Homologous recombination in cancer development, treatment and development of drug resistance. Carcinogenesis, 31, 955-960.

Homologous Recombination Antibody Portfolio

53BP1, Abraxas, ATM, ATR, ATRIP, BARD1, BLM, BRCA1, BRCA2, BRCC36, cAbl, Chk1, Chk2, Claspin, CtIP, FANCA, FANCD2, FANCE, H2AX, Mre11, NBS1, PALB2, Rad17, Rad50, RAD51C, Rad52, Rad9, RAP80, RFC1, RFC2, RFC3, RFC4, RFC5, SMC1, SMC3, SMC5, TopBP1, WRN.

Focusing on CML

2011-08-29T05:06:00.000-07:00

Chronic Myelogenous Leukemia (CML) is a hematopoietic stem-cell neoplasm of myeloid origin. A common method for the diagnosis of CML is based on the detection of a chromosomal translocation, first discovered in 1960 by Peter C. Nowell at the University Of Pennsylvania School Of Medicine in Philadelphia. The presence of the translocation was termed the Philadelphia chromosome and was the first chromosomal abnormality positively linked to cancer. Because it is present in over 90% of patients with CML, the Philadelphia chromosome is recognized as the genetic hallmark of CML. The translocation in the Philadelphia chromosome results in the juxtaposition of a portion of the BCR (breakpoint cluster region) gene located on chromosome 22 next to the ABL (Abelson leukemia virus) gene on chromosome 9 [t(9:22)]. Following the discovery of the Philadelphia chromosome came the finding that the product of the ABL gene function ed as a protein tyrosine kinase and that inappropriate regulation of kinase enzymes can be oncogenic. The BCR-ABL fusion protein was shown to be oncogenic and exhibit constitutive kinase activity. Constitutive kinase activity promotes the activation of a number of signaling pathways that support the uncontrolled growth and survival of hematopoietic cells resulting in neoplastic disease. These findings prompted a search for agents that could selectively target the activity of the BCR-Abl fusion protein and resulted in the identification of imatinib mesylate (Gleevec), the first tyrosine kinase inhibitor successfully and effectively used as a molecularly-targeted treatment for cancer. Thus, the discovery that began in Philadelphia with CML has served as a prime example of the successful translation of knowledge gained at the bench of basic scientists into treatments at the bedside of cancer patients.

Selected Reviews

Bumbea, H., Vladareanu, A. M., Voican, I., Cisleanu, D., Barsan, L., & Onisai, M. (2010). Chronic myeloid leukemia therapy in the era of tyrosine kinase inhibitors--the first molecular targeted treatment. J. Med. Life, 3, 162-166.

Koretzky, G. A. (2007). The legacy of the Philadelphia chromosome. J. Clin. Invest, 117, 2030-2032.

Sawyers, C. L. (1999). Chronic myeloid leukemia. N. Engl. J. Med., 340, 1330-1340.

CML Antibody Portfolio

Axl, BCR, cAbl, NUP98, SMARCB1/SNF5

Focusing on Ribosomes

2011-08-25T14:44:00.000-07:00

Ribosomes are large macromolecules that are constructed of both RNA and protein components and play a fundamental role in decoding genetic information. The RNA component of ribosomes has been shown to provide the enzymatic activities required for protein translation, while the protein components are thought to serve largely in a structural manner. The human 80S ribosome is made up of a 60S and 40S subunit. The 60S subunit is composed of a 5S rRNA, 5.8S rRNA, 28S rRNA and approximately 49 proteins and functions to catalyze peptide bond formation. The 40S subunit is composed of an 18S rRNA and approximately 33 proteins and functions to bring together the mRNA codon and the tRNA anticodon. Evolutionarily speaking, ribosomal proteins represent some of the oldest proteins, and their evolutionary development is considered to be the spark that facilitated a transition from a mainly DNA/RNA world to a DNA/RNA/protein wo rld. As a link between the RNA and protein world, ribosomes fulfill a critical element of the “central dogma” which hypothesizes that for all living things, DNA is transcribed into RNA which is then translated to protein. Ribosomal structure and function is highly conserved across archaea, bacteria, and eukaryotes and it is suggested that along with RNA and DNA, modern ribosomal macromolecules were present among the last common ancestor (LUCA). Because of their ancient beginnings and macromolecular complexity, studies of ribosome structure, function, and assembly will provide clues to the origins of life and the evolutionary history of modern species.

Selected Reviews

Crick, F. (1970). Central dogma of molecular biology. Nature, 227, 561-563.

Fox, G. E. (2010). Origin and evolution of the ribosome. Cold Spring Harb. Perspect. Biol., 2, a003483.

Moore, P. B. & Steitz, T. A. (2002). The involvement of RNA in ribosome function. Nature, 418, 229-235.

Ribosome Antibody Portfolio

AATF/Che-1, BOP1, DAP3, DjC21, Drosha, GCN1L1, GCN2, NPM1, NUFIP2, PRMT3, RACK1, RPL26, RPL7, rpL7a/SURF3, RPLP0, RPS6, RPS6 (S235), RPS6 (S235/S236), Ubiquitin.

Rabbit IL-8 (CXCL8) ELISA Kit

2011-07-18T05:55:00.000-07:00

Interleukin-8 (IL-8), renamed CXCL8, is a chemokine produced by macrophages and other cell types such as epithelial cells and endothelial cells. IL-8 can be secreted by any cell with toll-like receptors which are involved in the innate immune response. The most frequently studied receptors of IL-8 are the G protein coupled serpentine receptors CXCR1 (previously named IL-8 receptor α) and CXCR2 (previously named IL-8 receptor β). Both monomer and homodimer forms of IL-8 were reported as potent inducers of CXCR1 and CXCR2.

IL-8 is one of the first major mediators of the inflammatory response. The primary function of IL-8 is the induction of chemotaxis in its target cells (e.g. neutrophil granulocytes). Initially, macrophages phagocytose the antigen. Upon processing the antigen, macrophages release chemokines, such as IL-8, to signal other immune cells to come to the site of inflammation. IL-8 serves as a chemical signal that attracts neutrophils at the site of inflammation, and therefore is also known as neutrophil chemotactic factor.

While neutrophil granulocytes are the primary target cells of IL-8, there is a relative wide range of cells (endothelial cells, macrophages, mast cells, keratinocytes) responding to this chemokine, too. IL-8 is believed to play a role in the pathogenesis of bronchiolitis, a common respiratory tract disease caused by viral infection.

Standard range is 320 - 5 pg/ml

Rabbit IL-8 ELISA Kit Catalog No. E121-800

CCL2 ELISA Kits for Pig and Bovine

2011-06-21T05:11:00.000-07:00

Chemokine (C-C motif) ligand 2 (CCL2) is a small cytokine belonging to the CC chemokine family that is also known as monocyte chemotactic protein-1 (MCP-1). CCL2 recruits monocytes, memory T cells, and dendritic cells to sites of tissue injury and infection.1-2 This chemokine is produced as a protein precursor containing signal peptide of 23 amino acids and a mature peptide of 76 amino acids.3-4 It is a monomeric polypeptide, with a molecular weight of approximately 13kDa. The cell surface receptors that bind CCL2 are CCR2 and CCR4.5.

CCL2 is found at the site of tooth eruption and bone degradation. In the bone, CCL2 is expressed by mature osteoclasts and osteoblasts and is under the control of nuclear factor κB (NFκB). CCL2 causes the degranulation of basophils and mast cells, an effect potentiated by pre-treatment with IL-3 and other cytokines.6-7

Pig CCL2 ELISA Kit E101-800

Bovine CCL2 ELISA Kit E11-800

Background References

1. Carr MW, Roth SJ, Luther E, Rose SS, Springer TA (April 1994). "Monocyte chemoattractant protein 1 acts as a T-lymphocyte chemoattractant". Proc. Natl. Acad. Sci. U.S.A. 91 (9): 3652–6.

2. Xu LL, Warren MK, Rose WL, Gong W, Wang JM (01 September 1996). "Human recombinant monocyte chemotactic protein and other C-C chemokines bind and induce directional migration of dendritic cells in vitro". J. Leukoc. Biol. 60 (3): 365–71.

3. Yoshimura T, Yuhki N, Moore SK, Appella E, Lerman MI, Leonard EJ (February 1989). "Human monocyte chemoattractant protein-1 (MCP-1). Full-length cDNA cloning, expression in mitogen-stimulated blood mononuclear leukocytes, and sequence similarity to mouse competence gene JE". FEBS Lett. 244 (2): 487–93.

4. Furutani Y, Nomura H, Notake M, Oyamada Y, Fukui T, Yamada M, Larsen CG, Oppenheim JJ, Matsushima K (February 1989). "Cloning and sequencing of the cDNA for human monocyte chemotactic and activating factor (MCAF)". Biochem. Biophys. Res. Commun. 159 (1): 249–55.

5. Craig MJ, Loberg RD (December 2006). "CCL2 (Monocyte Chemoattractant Protein-1) in cancer bone metastases". Cancer Metastasis Rev. 25 (4): 611–9.

6. Conti P, Boucher W, Letourneau R, Feliciani C, Reale M, Barbacane RC, Vlagopoulos P, Bruneau G, Thibault J, Theoharides TC (November 1995). "Monocyte chemotactic protein-1 provokes mast cell aggregation and [3H]5HT release". Immunology 86 (3): 434–40.

7. Bischoff SC, Krieger M, Brunner T, Dahinden CA (May 1992). "Monocyte chemotactic protein 1 is a potent activator of human basophils". J. Exp. Med. 175 (5): 1271–5.

Chicken IL-16 ELISA Kit

2011-06-13T13:34:00.000-07:00

Interleukin 16 (IL-16) is a pleiotropic cytokine that has been characterized as a chemoattractant for certain immune cells expressing the cell surface molecule CD4, and has effects on mixed lymphocyte reaction and inhibition of HIV viral replication. Because this cytokine was discovered in 1982 for its T lymphocyte chemotactic activity, it was named “Lymphocyte Chemoattractant Factor (LCF)” until it was designated interleukin-16 in 1995. The product of this gene is generated as a precursor molecule, pro-IL-16, which is then processed to yield two functional proteins. IL-16 forms homotetramers and this structure is required for its bioactivity. The cytokine function is exclusively attributed to the secreted C-terminal peptide, while the N-terminal product may play a role in cell cycle control. Caspase 3 is reported to be involved in the proteolytic processing of this protein. IL-16 is released by a variety of immune (T cells, esinophils, and dendritic cells) and non-immune (fibroblasts, epithelial, and neurona) cells (Center et al., 1997; Cruikshank et al., 2000). In the non-diseased state, IL-16 mRNA is almost exclusively expressed on lymphatic tissue, and high levels in T cells. During inflammation, IL-16 is synthesized in a number of other tissues (Smith and Humphries, 2009). The sequence, structure, and function of IL-16 are highly conserved in all species examined. Thus far, all species of IL-16 tested induce similar bioactivities on CD4+ T cells. Chicken IL-6 has been cloned by Min et al. (2004) and shows 86% sequence identity to duct pro-IL-16, and 49-52% to various mammalian homologues. The recombinant chicken IL-16 showed chemoattractive activity to splenic lymphocytes.

Chicken IL-16 ELISA Kit E33-800

Representative Data

For Picture Perfect IP/Western Blots

2011-06-03T19:01:00.000-07:00

ReliaBLOT®

ReliaBLOT IP/Western Blot Reagents and Procedures provide an improved method for the detection of immunoprecipitated proteins assayed via Western Blot.

When the ReliaBLOT Reagents and Procedures are properly used, background from heavy chain should be reduced to less than 1% of that seen with standard procedures.

For use with antibodies raised in rabbits.

ReliaBLOT^® Procedure

Ordering Information:

WB120 - 20 miniblots

WB120T - 5 miniblots

Bovine Albumin ELISA kit

2011-05-16T08:56:00.000-07:00

Albumin is the most predominant protein in serum of domestic animals and man. It serves several functions including the binding and transporting of fatty acids, hormones, and metal ions, the maintenance of osmotic pressure and pH, and binding of exogenous toxins and products of lipid oxidation (Bertucci, 2002). Bovine and human albumins are carbohydrate-free monomeric proteins composed of three homologous domains (Peters, 1985 and Sugio, 1999). Over time, development of large-scale purification methods of bovine albumin have translated those functions into diagnostic, cell culture, and microbiological applications, (Mannuzza 2002) allowing for numerous biochemical applications including ELISAs (Enzyme-Linked Immunosorbent Assay), immunoblots, and immunohistochemistry. Bovine serum albumin is also used as a nutrient in cell and microbial culture.

The Bovine Albumin ELISA kit can be used for the detection of bovine albumin (BSA) in serum, plasma, milk, colostrum, cell culture supernatant and other biological samples containing traces of BSA. This kit contains sufficient components to quantitate Bovine Albumin in up to 40 samples, tested in duplicate. The standard range is 0.69 – 500 ng/ml.

Catalog Number E11-113

New Antibodies Released in April

2011-05-08T18:13:00.000-07:00

New Antibodies qualified for WB/IP: ANKS1A, Ankyrin 1, BACH1, DDX52, EBP1, EDD1, HEXIM2, Intersectin 1, Intersectin 2, LRRFIP1, MPP8, NAB1, Nulp1, p150glued, PDE8B, PEX14, RFX7, RPC3, SNAP190, SSRP1, SUV3, TIF-IA, TPP1, TR2, Tuba, ZC3H8, ZNF24, ZONAB.

New Antibodies qualified for IHC: Abraxas, ARID2, BCCIP, DDX27, HBO, IP3R3, JAK2, MASTL, MKL1, POP1, TOPORS.

IL-1 beta ELISA kits for Dog and Horse

2011-04-25T07:47:00.000-07:00

Interleukin-1 (IL-1) was one of the first cytokines ever described. Its initial discovery was as a factor that could induce fever, control lymphocytes, increase the number of bone marrow cells and cause degeneration of bone joints. At that time, IL-1 was known under several other names including endogenous pyrogen, lymphocyte activating factor, haemopoetin-1 and mononuclear cell factor. It was around 1984-1985 when scientists confirmed that IL-1 was actually composed of two distinct proteins, now called IL-1α and IL-1β.[1]

The original members of the IL-1 superfamily are IL-1α, IL-1β, and the IL-1 Receptor antagonist (IL-1RA). IL-1α and -β are pro-inflammatory cytokines involved in immune defense against infection. The IL-1RA is a molecule that competes for receptor binding with IL-1α and IL-1β, blocking their role in immune activation. Recent years have seen the addition of other molecules to the IL-1 superfamily including IL-18[2] and six more genes with structural homology to IL-1α, IL-1β or IL-1RA. These latter six members are named IL1F5, IL1F6, IL1F7, IL1F8, IL1F9, and IL1F10. In accord, IL-1α, IL-1β, and IL-1RA have been renamed IL1F1, IL1F2, and IL1F3, respectively.[3][4]

Both IL-1α and IL-1β are produced by macrophages, monocytes, fibroblasts and dendritic cells. They form an important part of the inflammatory response of the body against infection. These cytokines increase the expression of adhesion factors on endothelial cells to enable transmigration of leukocytes to sites of infection and re-set the hypothalamus thermoregulatory center, leading to an increased body temperature which expresses itself as fever. IL-1 is also important in the regulation of hematopoiesis. IL-1β production in peripheral tissue has also been associated with hyperalgesia (increased sensitivity to pain) associated with fever.[5]

For the most part, these two forms of IL-1 bind to the same cellular receptor. This receptor is composed of two related, but non-identical, subunits that transmit intracellular signals via a pathway that is mostly shared with certain other receptors. These include the Toll family of innate immune receptors and the receptor for IL-18.

Background References

1. Dinarello CA (1994). "The interleukin-1 family: 10 years of discovery". Faseb J. 8 (15): 1314–25.

2. Huising MO, Stet RJ, Savelkoul HF, Verburg-van Kemenade BM (2004). "The molecular evolution of the interleukin-1 family of cytokines; IL-18 in teleost fish". Dev. Comp. Immunol. 28 (5): 395–413.

3. Sims JE, Nicklin MJ, Bazan JF, Barton JL, Busfield SJ, Ford JE, Kastelein RA, Kumar S, Lin H, Mulero JJ, Pan J, Pan Y, Smith DE, Young PR (2001). "A new nomenclature for IL-1-family genes". Trends Immunol. 22 (10): 536–7.

4. Dunn E, Sims JE, Nicklin MJ, O'Neill LA (2001). "Annotating genes with potential roles in the immune system: six new members of the IL-1 family". Trends Immunol. 22 (10): 533–6.

5. Morgan MM, Clayton CC, Heinricher MM (2004). "Dissociation of hyperalgesia from fever following intracerebroventricular administration of interleukin-1beta in the rat". Brain Res. 1022 (1-2): 96–100.

Dog IL-1 beta ELISA Kit E44-800

Horse IL-1 beta ELISA Kit E77-805

E3 Ligases of the Ubiquitin Protease System

2011-04-17T18:05:00.000-07:00

The ubiquitin protease system involves a four-step enzyme cascade, which incorporates at least three enzymatic activities. The E1 ubiquitin-activating enzyme plays a role in the first step of the pathway and is responsible for delivering activated ubiquitin to the reaction. The E2 ubiquitin-conjugating enzyme receives the activated ubiquitin and transfers it to the target protein. Transfer of ubiquitin to the target is facilitated by the E3 ligase. The E3 ligase is responsible for dictating substrate specificity and bridging the activity of the ubiquitin-conjugating E2 enzyme with its target. The E3 ligases are a large and diverse group of proteins that exists as multi-protein complexes or as single proteins. E3 enzymes can be classified into at least 3 groups based on their structural domains. These domains include the HECT (homologous to E6-associated protein C-terminus), RING (really interesting new gene), or U-box (a modified RING motif without the full complement of Zn2+-binding ligands) domain. Because E3 ligases dictate target specificity, this group of enzymes is being considered as a target in the treatment of human diseases caused by defects in the ubiquitin protease system.

Selected Reviews

1. Lee J, and Zhou P. 2010. Cullins and Cancer. Genes Cancer 1 (7): 690-699.

2. Robinson PA, and Ardley HC. 2004. Ubiquitin-protein ligases. J. Cell Sci. 117 (Pt 22): 5191-5194.

3. Scheffner M, and Staub O. 2007. HECT E3s and human disease. BMC. Biochem. 8 Suppl 1: S6.

E3 Ligase Product Portfolio

APC1, APC4, APC5, APC7, BARD1, BMI1, BRCA1, BRCC36, CAND1, Cbl-b, CBX8, c-Cbl, CDC16/APC6, CDC23/APC8, COP1/RFWD2, Cul2, Cul3, Cul4a, Cul5, Cul7, DDB1, DTL/CDT2, DTX3L/BBAP, E4F1, EDD1, FBW7, FBXO11, FBXO31, FLRF/RNF41, Hakai, HECTD1, HERC1, HERC2, HERC4, Itch, Lasu1/Ureb1, MGRN1, MIB2/Skeletrophin, MKRN1, MKRN2, NEDD4L, NP95/UHRF1, NRIP, p600, PAM, PARC/H7-AP1, Rad18, RFWD3, Synoviolin, TOPORS, Triad1, TRIM33/TIF1gamma, TRIM37, TRIM41, TRIP12, ZNRF2.

IFN gamma ELISA Kits

2011-04-11T18:31:00.000-07:00

Interferon-gamma (IFN-γ) is a dimerized soluble cytokine that is the only member of the type II class of interferons.[1] IFN-γ is the hallmark cytokine of Th1 cells (whereas Th2 cells produce IL-4 and Th17 cells produce IL-17). NK cells and CD8+ cytotoxic T cells also produce IFN-γ. IFN-γ is produced predominantly by natural killer (NK) and natural killer T (NKT) cells as part of the innate immune response, and by CD4 and CD8 cytotoxic T lymphocyte (CTL) effector T cells once antigen-specific immunity develops.[2]

IFN-γ, or type II interferon, is a cytokine that is critical for innate and adaptive immunity against viral and intracellular bacterial infections and for tumor control. Aberrant IFN-γ expression is associated with a number of autoinflammatory and autoimmune diseases. The importance of IFN-γ in the immune system stems in part from its ability to inhibit viral replication directly, but, most important, derives from its immunostimulatory and immunomodulatory effects.

Cellular responses to IFN-γ are activated through its interaction with a heterodimeric receptor consisting of Interferon gamma receptor 1 (IFNGR1) and Interferon gamma receptor 2 (IFNGR2). IFN-γ binding to the receptor activates the JAK-STAT pathway. IFN-γ also binds to the glycosaminoglycan heparan sulfate (HS) at the cell surface. However, in contrast to many other heparan sulfate binding proteins, where binding promotes biological activity, the binding of IFN-γ to HS inhibits its biological activity.[3]

IFN-γ has antiviral, immunoregulatory, and anti-tumor properties.[4] It alters transcription in up to 30 genes producing a variety of physiological and cellular responses. Amongst the effects are:

Increases antigen presentation of macrophages.
Activates and increases lysosome activity in macrophages
Suppresses Th2 cell activity.
Causes normal cells to increase expression of class I MHC molecules.
Promotes adhesion and binding required for leukocyte migration.
Promotes NK cell activity.
Activates antigen presenting cells (APCs) and promotes Th1 differentiation by upregulating the transcription factor T-bet.
Activates inducible Nitric Oxide Synthase iNOS

Bethyl Laboratories' ELISA kits for IFN gamma:

Bovine IFN gamma ELISA Kit Catalog No. E11-805

Cat IFN gamme ELISA kit Catalog No. E22-801

Horse IFN gamma ELISA kit Catalog No. E77-806

Pig IFN gamma ELISA kit Catalog No. E101-806

Bovine VEGF-A ELISA Kit

2011-03-27T17:06:00.000-07:00

Vascular endothelial growth factor A (VEGF-A) is a glycosylated mitogen that specifically acts on endothelial cells and has various other effects including mediating increased vascular permeability, inducing angiogenesis, vasculogenesis and endothelial cell growth, promoting cell migration, and inhibiting apoptosis. Alternatively spliced transcript variants of VEGF-A, encoding either freely secreted or cell-associated isoforms, have been characterized.[1]
As its name implies, VEGF-A activity has been studied mostly on cells of the vascular endothelium, although it does have effects on a number of other cell types (e.g., monocytes/macrophages, neurons, cancer cells, kidney epithelial cells). In vitro, VEGF-A has been shown to stimulate endothelial cell mitogenesis and cell migration. VEGF-A is also a vasodilator and increases microvascular permeability and was originally referred to as vascular permeability factor.

In humans, elevated levels of this protein are linked to POEMS syndrome, also known as Crow-Fukase syndrome.[2] Mutations in this gene have been associated with proliferative and nonproliferative diabetic retinopathy.[3]

Bovine VEGF-A ELISA Kit Catalog No. E11-808

References

1. Mattei MG, Borg JP, Rosnet O, Marmé D, Birnbaum D (February 1996). "Assignment of vascular endothelial growth factor (VEGF) and placenta growth factor (PLGF) genes to human chromosome 6p12-p21 and 14q24-q31 regions, respectively". Genomics 32 (1): 168–9.

2. Dispenzieri A (November 2007). "POEMS syndrome". Blood Rev. 21 (6): 285–99.

3. Watanabe D, Suzuma K, Suzuma I, Ohashi H, Ojima T, Kurimoto M, Murakami T, Kimura T, Takagi H (March 2005). "Vitreous levels of angiopoietin 2 and vascular endothelial growth factor in patients with proliferative diabetic retinopathy". Am. J. Ophthalmol. 139 (3): 476–81.

Bovine TNF alpha ELISA Kit

2011-03-18T21:37:00.000-07:00

Tumor necrosis factor-alpha (TNF-α), also referred to as tumor necrosis factor (TNF), is a cytokine involved in systemic inflammation and is a member of a group of cytokines that stimulate the acute phase reaction. The primary role of TNF is in the regulation of immune cells. TNF is able to induce apoptotic cell death, to induce inflammation, and to inhibit tumorigenesis and viral replication. Dysregulation of TNF production has been implicated in a variety of human diseases, including major depression[1], Alzheimer's disease[2] and cancer.[3]
TNF was originally thought to be produced primarily by macrophages, but it is produced also by a broad variety of cell types including lymphoid cells, mast cells, endothelial cells, cardiac myocytes, adipose tissue, fibroblasts, and neuronal tissue. Large amounts of TNF are released in response to lipopolysaccharide, other bacterial products, and Interleukin-1 (IL-1).

Two receptors, TNF-R1 (TNF receptor type 1; CD120a; p55/60) and TNF-R2 (TNF receptor type 2; CD120b; p75/80), can be bound to by TNF. TNF-R1 is expressed in most tissues, and can be fully activated by both the membrane-bound and soluble trimeric forms of TNF, whereas TNF-R2 is found only in cells of the immune system, and respond to the membrane-bound form of the TNF homotrimer.

Upon contact with their ligand, TNF receptors undergo a conformational change, leading to the dissociation of an inhibitory protein from the intracellular death domain. This dissociation enables the adaptor protein TRADD (tumor necrosis factor receptor type 1-associated death domain) to bind to the death domain, serving as a platform for subsequent protein binding. Following TRADD binding, three pathways can be initiated including activaiton of NF-κB, activation of MAPK pathways, and induction of death signaling.[4][5]

The myriad and often-conflicting effects mediated by these pathways indicate the existence of extensive cross-talk. Other factors, such as cell type, concurrent stimulation with other cytokines, or the amount of reactive oxygen species (ROS) present can shift the balance in favor of one pathway or another. Such complicated signaling ensures that, whenever TNF is released, various cells with vastly diverse functions and conditions can all respond appropriately to inflammation.

Bovine TNF alpha ELISA Catalog No. E11-807

References
1. Dowlati Y, Herrmann N, Swardfager W, Liu H, Sham L, Reim EK, Lanctôt KL (2010). "A meta-analysis of cytokines in major depression". Biol Psychiatry 67 (5): 446-457.

2. Swardfager W, Lanctôt K, Rothenburg L, Wong A, Cappell J, Herrmann N (2010). "A meta-analysis of cytokines in Alzheimer's disease". Biol Psychiatry 68 (10): 930-941.

3. Locksley RM, Killeen N, Lenardo MJ (2001). "The TNF and TNF receptor superfamilies: integrating mammalian biology". Cell 104 (4): 487–501.

4. Wajant H, Pfizenmaier K, Scheurich P (2003). "Tumor necrosis factor signaling". Cell Death Differ. 10 (1): 45–65.

5. Chen G, Goeddel DV (2002). "TNF-R1 signaling: a beautiful pathway". Science 296 (5573): 1634–5.

6. Cerami A, Beutler B (1988). "The history, properties, and biological effects of cachectin". Biochemistry 27 (20): 7575–7582.

7. Vilcek J, Lee TH (1991). "Tumor necrosis factor. New insights into the molecular mechanisms of its multiple actions". J. Biol. Chem. 266 (12): 7313–7316.

IL-17A ELISA Kits

2011-03-07T18:33:00.000-08:00

Interleukin-17 (IL-17 or IL-17A) is the founding member of a group of cytokines called the IL-17 family. IL-17A was originally identified as a transcript from a rodent T-cell hybridoma by Rouvier et al. in 1993. Known as CTLA8 in rodents, IL-17 shows high homology to viral IL-17 encoded by an open reading frame of the T lymphotropic rhadinovirus Herpesvirus saimiri.[1] To elicit its functions, IL-17 binds to a type I cell surface receptor called IL-17R of which there are at least three variants IL17RA, IL17RB, and IL17RC.[2] In addition to IL-17A, members of the IL-17 family include IL-17B, IL-17C, IL-17D, IL-17E (also called IL-25), and IL-17F. All members of the IL-17 family have a similar protein structure, with four highly conserved cysteine residues critical to their 3-dimensional shape, yet they have no sequence similarity to any other known cytokines.

The most notable role of IL-17 is it involvement in inducing and mediating proinflammatory responses. IL-17 is commonly associated with allergic responses. IL-17 induces the production of many other cytokines (such as IL-6, G-CSF, GM-CSF, IL-1β, TGF-β, TNF-α), chemokines (including IL-8, GRO-α, and MCP-1), and prostaglandins (e.g., PGE2) from many cell types (fibroblasts, endothelial cells, epithelial cells, keratinocytes, and macrophages). The release of cytokines causes many functions, such as airway remodeling, a characteristic of IL-17 responses. The increased expression of chemokines attracts other cells including neutrophils but not eosinophils. IL-17 function is also essential to a subset of CD4+ T-Cells called T helper 17 (Th17) cells. As a result of these roles, the IL-17 family has been linked to many immune/autoimmune related diseases including rheumatoid arthritis, asthma, lupus, allograft rejection and anti-tumour immunity.[3]

Much progress has been made in the understanding of the regulation of IL-17. At first, Aggarwal et al. showed that production of IL-17 was dependent on IL-23.[4] Later, a Korean group discovered that STAT3 and NF-κB signalling pathways are required for this IL-23-mediated IL-17 production.[5] Consistent with this finding, Chen et al. showed that another molecule, SOCS3, plays an important role in IL-17 production.[6] In the absence of SOCS3, IL-23-induced STAT3 phosphorylation is enhanced, and phosphorylated STAT3 binds to the promotor regions of both IL-17A and IL-17F increasing their gene activity. In contrast, some scientists believe IL-17 induction is independent of IL-23. Several groups have identified ways to induce IL-17 production both in vitro[7] and in vivo[8][9] by distinct cytokines, called TGF-β and IL-6, without the need for IL-23.[7][8][9] Although IL-23 is not required for IL-17 expression in this situation, IL-23 may play a role in promoting survival and/or proliferation of the IL-17 producing T-cells. Recently, Ivanov et al. found that the thymus specific nuclear receptor, ROR-γ, directs differentiation of IL-17-producing T cells.[10]

Pig IL-17A ELISA Catalog No. E101-807

Bovine IL-17A ELISA Catalog No. E11-806

References
1. Rouvier E, Luciani MF, Mattéi MG, Denizot F, Golstein P (1993). "CTLA-8, cloned from an activated T cell, bearing AU-rich messenger RNA instability sequences, and homologous to a herpesvirus saimiri gene". J. Immunol. 150 (12): 5445–56.

2. Starnes T, Broxmeyer HE, Robertson MJ, Hromas R (2002). "Cutting edge: IL-17D, a novel member of the IL-17 family, stimulates cytokine production and inhibits hemopoiesis". J. Immunol. 169 (2): 642–6.

3. Aggarwal S, Gurney AL (2002). "IL-17: prototype member of an emerging cytokine family". J. Leukoc. Biol. 71 (1): 1–8.

4. Aggarwal S, Ghilardi N, Xie MH, de Sauvage FJ, Gurney AL (2003). "Interleukin-23 promotes a distinct CD4 T cell activation state characterized by the production of interleukin-17". J. Biol. Chem. 278 (3): 1910–4.

5. Cho ML, Kang JW, Moon YM, Nam HJ, Jhun JY, Heo SB, Jin HT, Min SY, Ju JH, Park KS, Cho YG, Yoon CH, Park SH, Sung YC, Kim HY (2006). "STAT3 and NF-kappaB signal pathway is required for IL-23-mediated IL-17 production in spontaneous arthritis animal model IL-1 receptor antagonist-deficient mice". J. Immunol. 176 (9): 5652–61.

6. Chen Z, Laurence A, Kanno Y, Pacher-Zavisin M, Zhu BM, Tato C, Yoshimura A, Hennighausen L, O'Shea JJ (2006). "Selective regulatory function of Socs3 in the formation of IL-17-secreting T cells". Proc. Natl. Acad. Sci. U.S.A. 103 (21): 8137–42.

7. Veldhoen M, Hocking RJ, Atkins CJ, Locksley RM, Stockinger B (2006). "TGFbeta in the context of an inflammatory cytokine milieu supports de novo differentiation of IL-17-producing T cells". Immunity 24 (2): 179–89.

8. Mangan PR, Harrington LE, O'Quinn DB, Helms WS, Bullard DC, Elson CO, Hatton RD, Wahl SM, Schoeb TR, Weaver CT (2006). "Transforming growth factor-beta induces development of the T(H)17 lineage". Nature 441 (7090): 231–4.

9. Bettelli E, Carrier Y, Gao W, Korn T, Strom TB, Oukka M, Weiner HL, Kuchroo VK (2006). "Reciprocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells". Nature 441 (7090): 235–8.

10. Ivanov, II, B.S. McKenzie, L. Zhou, C.E. Tadokoro, A. Lepelley, J.J. Lafaille, D.J. Cua, and D.R. Littman. 2006. The orphan nuclear receptor ROR-γ directs the differentiation program of proinflammatory IL-17+ T helper cells. Cell 126:1121-1133.

Anti-MAFs for IHC

2011-02-28T11:24:00.000-08:00

MAF family members are homologs of the musculoaponeurotic fibrosarcoma oncogene of the avian AS42 virus. MAF family members bear a well-conserved basic region and leucine zipper (b-Zip) motif that mediates DNA binding and dimer formation. The large MAF proteins, c-MAF, MAFB, and NRL also bear an acidic domain, a domain which functions in transcriptional activation (1). MAFs have been found to play key roles in development and have been especially studied in the development of the lens (2) and pancreas (3). Recently DeFalco et al from Duke University have further explored the developmental roles of MAFs and have discovered that MAFB and C-MAF also play a role in testis development (4). Because the Drosophila factor traffic jam (tj) has been shown to regulate gonad development in Drosophila (5), DeFalco et al set out to study the developmental expression of MAFs in the mouse testes during the embryonic stages of sexual differentiation. Using antibodies available from Bethyl Laboratories against MafB, MafA, and c-Maf in conjuction with Mafb-GFP transgenic mice, this group was able to characterize the localization of the MAF factors during gonadal development and analyze the origins of leydig cells which reside in the interstitial space. They report that MAFB, C-MAF, and VCAM qualify as markers of distinct cell-types that will prove to be tools useful for the understanding of gonad morphogenesis.

Antibodies against MAFs available from Bethyl Laboratories:

MafA, MafB, c-Maf

References

1. H. Motohashi, J. A. Shavit, K. Igarashi, M. Yamamoto, J. D. Engel, Nucleic Acids Res. 25, 2953 (1997).

2. H. M. Reza, K. Yasuda, Dev. Dyn. 229, 440 (2004).

3. M. E. Cerf, Eur. J. Endocrinol. 155, 671 (2006).

4. T. Defalco, S. Takahashi, B. Capel, Dev. Biol. (2011).

5. M. A. Li, J. D. Alls, R. M. Avancini, K. Koo, D. Godt, Nat. Cell Biol. 5, 994 (2003).

How do I love Thee…

2011-02-21T17:10:00.000-08:00

I’m not a romantic. T’is why I am late in my timing, one week post Valentine’s Day, to discuss the biological basis of love and mate-choice. You see how I say that, “the biological basis of love and mate choice”? Why is it that I must suggest that people don’t just find their true love with a gaze across a crowded room, but instead are driven by some pre-determined genetic factor that ties in with that whole survival-of-the-fittest story? …Because I am not a romantic? Maybe. But more likely because I am, after all, a biologist, and it is rare that I won’t try to find a biological or evolutionary explanation for most every social and cultural issue I contemplate. For the non-human animal world, the idea of genetic and evolutionary factors driving mate-choice is well established. I myself have known this since the ’70s, thanks to episodes of Wild Kingdom, which not only taught me that if you don’t have the genes that make you run fast, you’re gonna be someone’s dinner; but also, that if you DO have the genes that give you the biggest rack, the best dance, and the nicest tail (feathers) then you were gonna get taken out to dinner. I guess for humans we can also say that big racks play a role in mate choice along with other outward physical attributes, social status, and income earning potential (did I say I wasn‘t a romantic…ha!) . But what other animal instincts are still left with us that help us choose our mate? As humans we don’t really think about the olfactory cues that we unknowingly might send to our counterparts of the opposite sex; but through scientific studies this has been positively demonstrated by various studies including the “sweaty T-shirt experiments” (reviewed in (1)). Along with those experiments, numerous studies have shown that, like animals, humans have signature odors (reviewed in (2)), and those signature odors are related to the genotype of one’s MHC (major histocompatibility complex). The “sweaty T-shirt experiments” showed this at work in humans and revealed that females prefer the odor of T-shirts worn by males of a dissimilar genotype for the MHC. Additionally, there are other studies that reported spouses of a particular population were significantly MHC-dissimilar over random pairs of individuals (1). So why would one care about someone’s MHC? The MHC is a group of genes that encode proteins that play a role in self-recognition and are critical to immunity. They are glycoproteins expressed on the surface of nearly every cell type and function to display antigens that define a cell as “self”. Recognition of “self” is key to the innate properties of the immune system. By recognizing “self”, the immune system is kept from launching an immune response against one’s self; but in the event of infection, foreign peptides incorporated and displayed by MHC proteins can illicit an effective immune response. So, it is theorized that if a population of individuals exhibits a variety of MHC polymorphism, that might ensure that a population would be safeguarded against an epidemic infection (reviewed in (2)). What this means is that if your baby’s daddy (or momma) provides variety of MHC to your offspring, then there is a survival advantage! Now comes my attempt at being match-maker: How does one choose the best MHC? Firstly, listen to your nose, because it seems that these self-antigen presenting MHC proteins can be shed from cells, into the bloodstream, and to body fluids such as sweat, urine, or saliva (so that’s what kissing is all about!), making them available as odorants and attractants for olfactory analysis; secondly, choose an MHC-type different from your own so that your offspring receives alternate alleles that brings an advantageous variety to their immune system. Lastly, just when you decide, “He/She is not my type”, think again, because that just might be the cue you need to go for it. Fly to her side and make her your own or all thru your life you may dream all alone, and once you have found her, never let her go!

References

1. R. Chaix, C. Cao, P. Donnelly, PLoS. Genet. 4, e1000184 (2008).

2. P. B. Singh, Reproduction. 121, 529 (2001).

Antibodies available from Bethyl Laboratories against proteins important to immunity:

ATF2, BMI1, C/EBP beta, C1QBP, CAR, CARMA1, Cbl-b, CERT/GPBP, Coronin 1, CREB, E4BP4, ERK1, ERK2, FAS, FOXP3, HSP60, I kappa B-beta, IKK-alpha, IKK-beta, ICAM-1, IRF3, Itch, MALT1, MAVS/VISA, MEK1, MEK2, MEKK1, MEKK2, NF-kappaB1, NF-kappaB2, Notch1, p38 MAPK, PACT

Bovine CCL2, Bovine CXCL9, Bovine IFN-gamma, Bovine IL-13, Bovine IL-2, Cat IL-2, Chicken IL-16, Dog IL-1 beta, Dog IL-2, Horse CCL2, Horse IL-1 beta, Horse IL-2, Horse IL-4, Pig CCL2, Pig CXCL10, Pig IFN-beta, Pig IFN-gamma, Pig IL-13, Pig IL-17A, Pig IL-8

Bethyl Laboratories: Excellence in IHC

2011-02-14T06:16:00.000-08:00

Bethyl Laboratories offers a line of high quality polyclonal antibodies specifically validated for IHC.

The following abstract was published in the April 2009 FASEB Journal Meeting Abstract Supplement (1006.8). The abstract describes the superior utility of Bethyl Laboratories Rabbit Anti-Human CD31 (#IHC-00055) for staining vasculature in fixed tumor tissue:

Evaluation of vascular endothelial staining by various antibodies in human and xenograft tumors using three different methods of fixation

Ann E Breckenkamp1, Karen Mintze1, Versie Barnes1, Tasha Vance1, Nicholas Pierson1, Jenna Pangallo1 and George Sandusky1

1 Pathology, Indiana University School of Med, Indianapolis, IN

ABSTRACT

Light microscopy was used to compare the staining of various antibodies against the vascular endothelial markers CD31, CD34, CD105, EphB2, and EphB4 in human and xenograft tissues that were fixed in three different fixatives: 4% paraformaldehyde (PF), 10% Neutral Buffered Formalin (NBF), or Zinc-Tris (ZT). Since the ability of an antibody to bind to its target molecule depends on the fixation method, it would be useful to identify antibodies that stain tumor vasculature regardless of which fixative was used to preserve the tissue. Each antibody evaluated was optimized in ZT fixed tissues, and the protocol was used to stain all samples, except for two antibodies that did not stain ZT fixed tissues. Bethyl Laboratories Rabbit Anti-Human CD31 (#IHC-00055) and Cell Signaling Mouse Anti-Human CD34 (#3569) gave excellent staining in human tissues prepared in 4% PF, 10% NBF, and ZT, in contrast to the other antibodies evaluated. The Bethyl antibody also gave moderate staining in HCT116 xenografts prepared in all three fixatives, making it the only antibody evaluated that was useful for staining vasculature in human breast, colon, and lung tissues and xenografts for all three fixatives.

Detection of Human CD31 in FFPE section
of human colon adenocarcinoma.

IHC Simplified

2011-02-05T17:28:00.000-08:00

Do you want to produce beautiful IHC images for your publications? If yes, then try the IHC Accessory Kit from Bethyl Laboratories and see how much it can simplify your life in the lab. If you want a quick, reliable, proof-of-concept kit to jump-start your IHC needs then this kit is for you! Bethyl Laboratories IHC Accessory Kit is perfect way to introduce beginners to IHC methodology with easy-to-follow step-by-step instructions. All the reagents you need are included in the kit, and when used in conjunction with Bethyl’s line of IHC validated primary antibodies you will be well on your way to producing beautiful immunostaining with a minimal investment of your time, effort, and valuable research dollars.

Immunoprecipitation for the Analysis of snRNP Formation

2011-01-31T05:15:00.000-08:00

snRNPs (small nuclear ribonucleoproteins) are complexes of protein and RNA that play a critical role in the splicing and removal of introns of pre-mRNAs. There are at least five major snRNPs: U1, U2, U4, U5, and U6, and each associate with their own small nuclear RNA (snRNA) component: U1snRNA, U2snRNA, U4snRNA, U5snRNA, and U6snRNA. The assembly of snRNPs and splicing complexes is intricate and highly regulated. The survival of motor neuron (SMN) protein has been shown to be essential to the assembly of splicesomal snRNPs and Cajal body formation. SMN deficiency is associated with spinal muscular atrophy (SMA), and defects in snRNP biogenesis have been found to be a cause of SMA. Recently Boulisfane et al set out to study the effects of SMN mutations on the formation of complexes involved in nuclear pre-mRNA splicing. To investigate snRNP formation and composition, the group used antibodies against components of snRNP particles to immunoprecipitates (IP) snRNPs from SMA cell lysates. RNA was extracted from the associated immunocomplexes and the RNA analyzed by northern blot using complementary snRNA probes. Immunoprecipitation experiments were performed with antibodies against the 110K tri-snRNP-specific protein, the 100K and 40K components of U5 and tri-snRNPs, Prp5- a component of the 17S U2 snRNP, and SF3b155- a component of the U2 snRNP. Anti-100K/DDX23, anti-110K/SART1, anti-Prp5/DDX46 and anti-SF3b155 were purchased from Bethyl Laboratories. Via this application, it was revealed that SMA cells display differential splicing defects, Cajal body disruption, and snRNP assembly due to SMN deficiency.

Reference:

Boulisfane, N., Choleza, M., Rage, F., Neel, H., Soret, J., and Borndonne, R. (2011). Impaired minor tri-snRNP assembly generates differential splicing defects of U12-type introns in lymphoblasts derived from a type I SMA patient. Hum. Mol. Genet. 20 (4): 641-648.

Bethyl Laboratories Antibodies Used To Study the Role of PHF8 in Cell Cycle Control

2011-01-17T05:27:00.000-08:00

With the help of a series of antibodies from Bethyl Laboratories, Wen Liu et al from the University of San Diego have demonstrated that PHF8 can function as a demethylase for histone H4 lysine 20 and plays a key role in the loading of the condensin II complex and regulation of the cell cycle (Liu et al., 2010). The PHF8 protein bears two functional domains, a PHD (plant homeodomain) finger and a JmjC (Jumonji-like C-terminus) domain. The gene for PHF8 is located on the X-chromosome (Xp11), and mutations in the PHF8 gene are associated with X-linked mental retardation (XLMR) and cleft lip/cleft palate (Laumonnier et al., 2005). From studies of its structure, PHF8 had been proposed to play a role in transcriptional regulation and histone demethylation. The results, published by Liu et al in the July 22, 2010 issue of Nature, have confirmed a role for PHF8 in the demethylation of H3K9me1/2, H3K27me2, and H4K20me1. The study has also shown that PHF8 plays a role in G1-S transition by binding and regulating the promoters of cell cycle genes and by dissociating from chromatin in early mitosis, an event associated with an accumulation of H4K20me1. The association between mutations in demethylase genes such as PHF8 and human disease indicates an importance for the further understanding of this class of enzymes.

The Bethyl Products used in this study included:

Target	Cat. No.	Application
CAP-D2	A300-601A	western blot
CAP-G	A300-602A	western blot
CAP-H	A300-603A	western blot
CAP-H2	A302-275A	western blot
HCF1	A301-399A	western blot
PHF8	A301-772A	western blot
PHF8	IHC-00343	IHC
SET1A	A300-290A	western blot
SMC2	A300-058A	IF
SMC4	A300-064A	ChIP

References

Laumonnier, F., Holbert, S., Ronce, N., Faravelli, F., Lenzner, S., Schwartz, C. E. et al. (2005). Mutations in PHF8 are associated with X linked mental retardation and cleft lip/cleft palate. J Med.Genet., 42, 780-786.

Liu, W., Tanasa, B., Tyurina, O. V., Zhou, T. Y., Gassmann, R., Liu, W. T. et al. (2010). PHF8 mediates histone H4 lysine 20 demethylation events involved in cell cycle progression. Nature., 466, 508-512.

New Antibodies and Kits Released in December

2011-01-13T14:02:00.000-08:00

New antibodies qualified for WB/IP: ALK, ANKLE2, c-Jun, DGKzeta, Frizzled-6, RNF34, BE2Q2,
Zwilch.

New antibodies qualified for IHC: ABC50, ANKRD17, BCAR1/p130CAS, CKII beta, CROP/Luc7A, Cyclin C, eIF3A/eIF3S10, GBF1, PDZ-GEF1, PRKAR2A/PKA-RIIalpha, PRPF4B, RanGAP1, Shc1,
UVRAG, ZAK.

New ELISA kits for: Chicken IgA, Chicken IgG, Chicken IgM, Human CRP, Mouse IgG2c, Rat Albumin, Rat IgA, Rat IgG, Rat IgM.

TRIM16 Found to Act as a Tumor Suppressor

2011-01-13T12:37:00.000-08:00

A recent report in Oncogene [Oncogene (2010) 29, 6172–6183] demonstrates that TRIM16 can act as a tumor suppressor in neuroblastoma cells. TRIM16 was originally identified as a gene termed EBBP (estrogen-responsive B box protein) and reported as a gene up-regulated by estrogen and tamoxifen in human mammary epithelial cells (Liu et al., 1998) and as a gene regulated by keratinocyte growth factor in human keratinocytes (Beer et al., 2002). EBBP was also found to play a role in in innate immunity as a binding partner of IL-1beta (Munding et al., 2006), and later TRIM16 was shown to be a DNA-binding protein that possessed histone acetyltransferase activity and to mediate growth inhibition of retinoid-treated cancer cells (Cheung et al., 2006; Raif et al., 2009). Together, these findings suggest that TRIM16 might function as a potential target for anti-cancer therapies. The most recent report on TRIM16 shows that overexpression of TRIM16 can reduce growth of neuroblastoma cells, enhance retinoid-induced differentiation, and reduce tumorigenicity in vivo. The study also demonstrates that TRIM16 accomplishes its tumor suppressor activities via its interactions with cytoplasmic vimentin and nuclear E2F1. This study strengthens the proposal that TRIM16 is a viable target for the development of ant-cancer therapies (Marshall et al., 2010).

Bethyl Laboratories antibodies available for the study of TRIM16, Vimentin, and E2F1
Reference List

Beer, H. D., Munding, C., Dubois, N., Mamie, C., Hohl, D., & Werner, S. (2002). The estrogen-responsive B box protein: a novel regulator of keratinocyte differentiation. J Biol Chem., 277, 20740-20749.
Cheung, B. B., Bell, J., Raif, A., Bohlken, A., Yan, J., Roediger, B. et al. (2006). The estrogen-responsive B box protein is a novel regulator of the retinoid signal. J Biol Chem., 281, 18246-18256.

Liu, H. L., Golder-Novoselsky, E., Seto, M. H., Webster, L., McClary, J., & Zajchowski, D. A. (1998). The novel estrogen-responsive B-box protein (EBBP) gene is tamoxifen-regulated in cells expressing an estrogen receptor DNA-binding domain mutant. Mol Endocrinol., 12, 1733-1748.

mTOR Signaling, Cilia, and Polycystic Kidney Disease

2011-01-09T12:36:00.000-08:00

Evidence has emerged for a role for mTOR in the genesis of autosomal dominant polycystic kidney disease (ADPKD), a genetic disease characterized by the formation of renal cysts [reviewed in (Boletta, 2009) and (Weimbs, 2007)]. ADPKD has been linked to mutations in the genes polycystin-1 (PC-1) and polycystin-2 (PC-2). PC-1 and PC-2 form a membrane complex localized to cell-cell junctions, renal cilia, and urinary exosomes. The signal transuducing activity of the PC-1/PC-2 complex is believed to be essential to prevent renal cystogenesis, but the mechanisms involved are not positively known. From several studies it appears that the mTOR pathway is disregulated in polycystic kidney disease and that PC-1 may regulate the mTOR pathway to influence cell growth and cyst formation. Additionally, since the PC-1/PC-2 complex localizes to primary cilia, it has been proposed that loss of cilia function can lead to mechanosensing defects and loss of growth control leading to cyst formation. These observations have led a group to investigate a possible link between cilia function and the regulation of cell size via the mTOR pathway (Boehlke et al., 2010). In this study, the authors made a positive connection between cilia function and the mTOR pathway and have shown that the mechanical stimulation of primary cilia downregulates mTOR signaling. These findings have now provided more clues to the mechanisms involved in the genesis of polycystic kidney disease and the possibilities of treatments that target the mTOR pathway.

References

Boehlke, C., Kotsis, F., Patel, V., Braeg, S., Voelker, H., Bredt, S. et al. (2010). Primary cilia regulate mTORC1 activity and cell size through Lkb1. Nat Cell Biol., 12, 1115-1122.

Boletta, A. (2009). Emerging evidence of a link between the polycystins and the mTOR pathways. Pathogenetics., 2, 6.

Weimbs, T. (2007). Polycystic kidney disease and renal injury repair: common pathways, fluid flow, and the function of polycystin-1. Am J Physiol Renal Physiol., 293, F1423-F1432.

Antibodies Available from Bethyl Laboratories for the study of polycystin and mTOR signaling:

Polycystin-2, 4EBP1, AKT1, AKT2, AktS1/PRAS40, AMPK alpha 1, AMPK alpha 2, BRAF, eIF4B, eIF4E, ERK1, ERK2, GbetaL, HIF1-alpha, MEK1, MEK2, mTOR, p70S6K, PDK1, PKC-alpha, PKC-delta, PTEN, RAF1/c-RAF, Raptor, REDD1, Rictor, RPS6, RSK1, RSK2, Sin1, TSC1, TSC2.

The Rhythms of Methyl Transfer and Alternative Splicing

2011-01-03T13:05:00.000-08:00

In the Nov 4th 2010 issue of Nature, Sanchez et al report a molecular mechanism that links the circadian clock to the control of RNA processing. They found that mutations in PRMT5 impair circadian rhythms in both an Arabidiopsis Thaliana and Drosophila melanogaster model. The impairments are caused by alterations in mRNA splicing of core-clock genes, and the findings demonstrate a positive association between the expression profile of PRMT5, the regulation of alternative splicing, and the synchronization of the circadian clock. (Sanchez et al., 2010)

References

Sanchez, S. E., Petrillo, E., Beckwith, E. J., Zhang, X., Rugnone, M. L., Hernando, C. E. et al. (2010). A methyl transferase links the circadian clock to the regulation of alternative splicing. Nature., 468, 112-116.

Bethyl Laboratories' Portfolio of Antibodies

PRMT5, BMAL1, CKI delta, CLOCK, Cry1, Cry2, Dec1, Timeless

New Antibodies Released in November

2010-12-13T18:02:00.000-08:00

New antibodies qualified for WB/IP: ALIX, Daple, Dishevelled 2, NIPA, PGRMC2, PLC-gamma 1, Sgt1 (SUGT), Synoviolin, UCH-L3, WWP1.

New antibodies qualified for IHC: ARHGEF16, Beta-catenin, eIF2A, KSRP, Menin, NF-YA, NF-YC, PDZ-RhoGEF, Prohibitin, SAM68, vPARP.

New Antibodies Released in October

2010-11-16T10:41:00.000-08:00

New antibodies qualified for WB/IP: Aly, Atg4B, AUP1, CacyBP, CAND1, Cbl-b, DUBA, Gigaxonin, HECTD1, JAM-A, KLHL12, MGRN1, NFX1, OTUB1, RABGEF1, SAE1, SAE2, SENP1, Triad1, UBXD1, USP8, VAPB, VCIP135, WWP2.

New antibodies qualified for IHC: Diaphanous 1, eIF4G1/eIF4GI, EML4, FHL2, GEF-H1, NPPL1/SHIP2, Leo1, Menin, PKD2, QKI, TORC2.

Antibodies against Cyclins and Cyclin-Dependent Kinases

2010-11-01T09:36:00.000-07:00

Detection of CDK11 in FFPE section of
human stomach adenocarcinoma

Cyclin dependent kinases (CDKs) and their associated cyclin partners have long been recognized as complexes that play an integral role in the progression of the cell cycle. Recently, the well established role of the CDK/cyclin complexes has been broadened, and current studies show that an atypical group of CDK/cyclin complexes are also directly involved in activities such as transcription, translation, and mRNA processing. The transcriptional regulating cyclins include cyclin C, H, K, L1, L2, T1, and T2 which associate with CDK8, -7, -9, and -11; the complexes formed by these factors appear to be involved in distinct aspects of transcription and splicing:

• Cyclin C associates with CDK8. CDK8/cyclin C has been identified as a component of the Mediator complex, a coactivator complex that provides a scaffold to link gene-specific transcription factors to RNA polymerase II and the general transcription machinery.

• Cyclin H forms a complex with CDK7and MAT1. The CDK7/cylin H/MAT1 complex has been identified as a component of the TFIIH multi-subuint general transcription factor. In this transcription complex, CDK7 is responsible for the phosphorylation of several components of the transcriptional machinery.

• Cyclin L1 and L2 associate with CDK11. CDK/cyclin L complexes have been implicated to play a role in pre-mRNA splicing.

• Cyclins T1, T2a, T2b and K form a complex with CDK9. CDK9 has been identified as a subunit of the transcription elongation factor p-TEFb. pTEFb is responsible for phosphorylating the carboxyl-terminal domain (CTD) of RNA polymerase II to facilitate elongation.

The discovery of novel roles for CDK/cyclin complexes provides an avenue to connect the processes of cell cycle progression, transcription, and RNA splicing into a common regulatory network and raises many questions concerning how these complexes achieve cross-talk to promote the complex coordination of cell cycle progression and gene expression (reviewed in Loyer, Trembley, Katona, Kidd, & Lahti, 2005).

Reference List
Loyer, P., Trembley, J. H., Katona, R., Kidd, V. J., & Lahti, J. M. (2005). Role of CDK/cyclin complexes in transcription and RNA splicing. Cell Signal., 17, 1033-1051.

Bethyl Laboratories' Portfolio of Antibodies against Cyclins and Cyclin-Dependent Kinases:

CDK2, CDK11, CDK5RAP2, CDK5RAP3, CDK7, CDK8, Cyclin C, Cyclin E1, Cyclin H, Cyclin K, Cyclin L1, Cyclin L2, Cyclin T2, Cyclin Y.

Antibodies to the CHD Proteins

2010-10-25T19:05:00.000-07:00

Detection of Human and Mouse
CHD4/Mi2beta by WB (h&m) and IP (h).

The process of chromatin remodeling is central to the regulation of DNA-associated activities such as replication, recombination, repair, and transcription. Chromatin remodeling involves the modulation of nucleosomal structures that restricts and regulates the access of DNA-binding factors to regulatory elements to direct nuclear activities. The modulation of nucleosomes and remodeling of chromatin is achieved through the ATP-dependent actions of multi-subunit remodeling complexes that alter chromatin structure through histone displacement and nucleosomal sliding. On the basis of their associated ATPase subunit, three families of chromatin-remodeling complexes have been identified: SWI2/SNF2-based complexes, ISWI-based complexes, and CHD (chromodomain-helicase-DNA-binding protein)-based complexes. Although the three families of complexes share similar structural and functional characteristics, they also possess their own unique properties.

The CHD family of proteins appears to be primarily involved in the regulation of developmental gene expression and heterochromatin formation. There are nine members of the CHD family of proteins (CHD1-9). The CHD family members possess 3 common structural and functional domains: a chromodomain (chromatin organization modifier), an SNF2-like helicase/ATPase domain, and a C-terminal DNA-binding domain. The chromodomain functions to recognize and bind nucleosomes while the SNF2-like helicase/ATP domain provides the energy required for histone displacement and nucleosomal sliding for remodeling. The DNA-binding domain is more divergent among the CHD family members and may partially function to promote sequence specific interactions with DNA. Despite their structural similarity, the functions of the CHD family members are diverse. Functional diversity is likely imparted by the divergence in the DNA binding domain, as well as differences in binding partners and expression patterns of the CHD family members. Continued study of the functional contribution of each CHD family member to chromatin remodeling will further our understanding of how appropriate gene activation and repression is achieved in the developing organism.

Bethyl Laboratories' Portfolio of Antibodies to CHD Proteins includes:

CHD1, CHD3, CHD4/Mi2 beta, CHD6, CHD7, CHD8, CHD9/CReMM.

Antibodies to General Transcription Factors

2010-10-20T07:04:00.000-07:00

Detection of Human and Mouse
GTF3C5/TFIIIC63 by Western Blot (h & m)
and Immunoprecipitation (h).

The transcription of genes involves a multitude of proteins that assemble on specific gene promoter elements for the synthesis of various types of RNA. In eukaryotes, transcription is carried out by three classes of RNA polymerases (pols): pol I, pol II, and pol III. Each RNA polymerase is responsible for the synthesis of a particular type of RNA. Pol I is involved in the synthesis of ribosomal RNAs; Pol II synthesizes mRNAs, snRNAs, and micro RNAs; while pol III synthesizes tRNAs, and other small RNAs. Pol II and III associate with a specific set of general transcription factors (GTFs) for the initiation of transcription. The GTFs are aptly termed for the RNA polymerase with which they associate. Pol II associates with a core transcription complex made up of six general transcription factors (GTFs). The six GTFs required for transcriptional initiation by RNA pol II include TFIIA, TFIIB, TFIID, TFIIE, TFIIF, and TFIIH. Additional pol II-associated factors involved in basal transcription have been identified and include TFIIG, TFII-I, TFIIJ, and TFIIK. Many of the GTFs for RNA pol II exist as multisubunit complexes. For example, TFIID is a complex of TATA-binding protein (TBP) and nine TBP-associated factors (TAFs). TFIIA, TFIIE, TFIIF, and TFIIH also exist as multisubunit complexes. TBP is the most well characterized subunit of TFIID and is also critical to transcription initiation at pol I and pol III promoters. At pol III promoters, TBP, along with the pol III GTFs, TFIIIA, TFIIIB, and TFIIC, recruit RNA pol III to core promoter elements. TFIIIB and TFIIIC also exist as multisubunit complexes. TFIIIB is made up of three subunits: hTBP, hBRF, and hB. TFIIIC is composed of 6 subunits: TFIIIC63, TFIIIC220, TFIIIC102, TFIIIC110, TFIIIC90, and TFIIIC35.

Selected Reviews
1. G. Orphanides, T. Lagrange, D. Reinberg, Genes Dev. 10, 2657-2683 (1996).
2. Y. Huang and R. J. Maraia, Nucleic Acids Res. 29, 2675-2690 (2001).

Bethyl Laboratories' Porfolio of Antibodies to General Transcription Factors includes

GTF2A1/TFIIA

GTF2B/TFIIB

GTF2E1/TFIIE-alpha

GTF2I/TFII-I

GTF3C1/TFIIIC220

GTF3C2/TFIIIC110

GTF3C3/TFIIIC102

GTF3C4/TFIIIC90

GTF3C5/TFIIIC63

GTF2IRD/TFII-IRD1

TFIIS

Antibodies to the CSN Complex

2010-10-13T12:43:00.000-07:00

The CSN (COP 9 signalsome) complex was originally identified as a regulator of light-mediated development in Arabidopsis that, when mutated, caused signal-independent expression of light-induced genes (thus COP, constitutive morphogenesis). In mammals, the CSN has been found to play a significant role in cellular processes such as cell signaling, cell cycle progression, gene transcription, cell survival, and DNA repair. Current findings show that the CSN complex functions as an interface between cellular signals and the regulation of ubiquitin-dependent degradation of proteins. The CSN appears to accomplish its functions through its metalloproteinase activity which is able to cleave Nedd8 from cullin-based E3-ubiquitin ligase (Cul-E3) as well as through its associated kinase activity. The mammalian CSN is comprised of eight subunits, CSN1 through CSN8. The CSN and its individual subunits have been observed to interact with a multitude of proteins, but the role of each subunit as well as the physiological relevance of these interactions is unclear. The ubiquitin-proteasome pathway and the CSN have been implicated in the development of human disease and cancer and are thus being considered as potential targets for therapeutic intervention. A better understanding of the roles of the individual CSN subunits, the relevance of interacting proteins, and the enzymatic activities of the CSN will allow a better evaluation of the value of the CSN as a therapeutic target.

CSN1 Antibody, CSN2 Antibody, CSN3 Antibody, CSN4 Antibody, CSN5 Antibody, CSN7b Antibody

New Polyclonal Antibodies released in September

2010-10-11T05:43:00.000-07:00

New antibodies qualified for WB/IP: C1QBP, Fas, gp78, ICAM-1, L3MBTL2, MAX, MGA, MUPP1, NS1-BP, RAD23A, Reticulon-3, RNF2, RNGTT, RPLP0, Sequestosome-1, SmcHD1, SND1, TFIIB, Ubinuclein-1, UNG.

New antibodies qualified for IHC/ICC: Caf1p150, KIAA0528, LARP1, PA1, PABP4, REDD1, THOC5, Ymer.

New Cytokine ELISA Kits - September

2010-10-07T18:55:00.000-07:00

Bovine CXCL9 ELISA Kit

Bovine IFN-gamma ELISA Kit

Bovine IL-13 ELISA Kit

Dog IL-2 ELISA Kit

Pig IFN-gamma ELISA Kit

Pig IL-17A ELISA Kit

Pig IL-8 ELISA Kit

New Polyclonal Antibodies Released in August

2010-10-01T08:01:00.000-07:00

New antibodies qualified for WB/IP: ATXN7L3, BAG3, BEGAIN, BNIP3L, CAR, CIAPIN1, DAP3, Derlin-1, ELF2, FAF1, FBP17, FYCO1, HJURP, HSP60, Huntingtin, Jade-1, KIAA0802, KNL-2, L3MBTL3, MAP1S, MgcRacGAP, NOL10, PolA1, PQBP1, PTPIP51, PUF60, RABEP1, RCAS1, RN-tre, SAP30BP, SLTM, SMG5, SPF30, SUPT7L, THOC1, TRAF4.

New antibodies qualifiied for IHC/ICC: ABC2, BIG2/ARFGEF2, COX4, CPSF100, FOXM1, FOXP3, GIT2, NP95/UHRF1, OSR1, RBM10, REDD1, SRcyp, U2AF35, ZNF592.

New Polyclonal Antibodies Released in July

2010-08-09T14:51:00.000-07:00

New antibodies qualified for WB/IP: AIF, AP4, API5, ARNT, Arpc1b, Bin1, C/EBP beta, GMEB1, Importin 4, Importin 7, MKL2, MSK1, MSK2, NFAT3, NOL12, NPAT, ORC2, ORC3, PDE3A, PDE3B, PDE4D, PRP6, PSMA3, PSMA5, PSMD11, Pygo2, RAP74, SGT1, STAT1, TAB2, TDP2, TFIIA, TFIIE-alpha, USP13, YY1.

New antibodies qualified for IHC: ANKRD28/PITK, C1orf55, CSTF2T/TauCSTF64, CTNND1, EEA1, ELF1, ERCC5/XPG, FALZ/BPTF, GIT1, JHDM1A, LAP1B, MAP4, MAP4K4/HGK, MAP7, NF-kappaB1, NF-kappaB2, Notch1, Notch2, Phospho RelA (S468), Phospho RelA (S536), RelA, SFRS2IP, Symplekin, WDR79.

Antibodies Against Forkhead Box (FOX) Proteins

2010-06-29T06:01:00.000-07:00

The first fork head (FKH) box (Fox) family member was discovered as a gene responsible for the "fork head" phenotype in D. melanogaster mutants. In these mutants, defects in FKH caused homeotic transformations that involved the replacement of foregut and hindgut by ectopic head structures resulting in two "spiked-head" structures in embryos (1). Since the discovery of FKH in flies, hundreds of Fox gene family members have been discovered in a range of species from yeast to humans. At least 43 members of the of the human Fox gene family have been identified (2); and based on phylogenetic analysis, 19 classes of Fox proteins have been established (FoxA-FoxS) (3). Fox proteins are defined by a highly conserved 100-residue forkhead DNA binding domain which consists of three alpha-helices, three-beta sheets, and two "wing" regions. The Fox proteins are DNA-binding proteins and generally function as transcriptional regulators; however duplication of the FKH domain through evolution has resulted in a family of proteins that exhibit a wide spectrum of functional diversity. The requirement of this family of proteins in processes ranging from organogenesis to speech acquisition exemplifies its importance through evolution, and along with its involvement in human disease, makes the Fox family a subject worthy of future study.

Selected Reviews
1. D. Weigel et al., "The Homeotic Gene Fork Head Encodes a Nuclear Protein and Is Expressed in the Terminal Regions of the Drosophila Embryo," Cell. 57, no. 4 (1989): 645-658.
2. M. Katoh and M. Katoh, "Human FOX Gene Family (Review)," Int.J Oncol. 25, no. 5 (2004): 1495-1500.
3. S. Hannenhalli and K. H. Kaestner, "The Evolution of Fox Genes and Their Role in Development and Disease," Nat Rev Genet. 10, no. 4 (2009): 233-240.

FOXC2
FOXK1
FOXK2
FOXM1
FOXO1a
FOXO3a
FOXO4
FOXP1
FOXP3
FOXP4

Antibodies to Proteins Involved in Histone Acetylation and Deacetylation

2010-06-23T06:25:00.000-07:00

Histone modification is central to chromatin architecture and the modulation of transcription. Histone deacetylases (HDACs) and histone acetylases (HATs) are chromatin-modifiying enzymes that catalyze the removal and addition of acetyl groups from acetyl-lysine residues on the N-terminal extensions of core histones. Acetylated histones are a key feature of actively transcribed genes. Histone acetylation and transcriptional activation is counteracted by deacetylation which results in transcriptional repression. Aberrant histone acetylation has been observed in human cancers. Recently it has become recognized that inhibiting chromatin-modifying enzymes such as HDACs may provide an avenue for the development of anti-cancer agents. Although this avenue has promise, the development of such agents still requires a more detailed analysis and better understanding of the complex regulatory mechanisms of histone acetylation.

Selected Reviews
1. P. Gallinari et al., "HDACs, Histone Deacetylation and Gene Transcription: From Molecular Biology to Cancer Therapeutics," Cell Res. 17, no. 3 (2007): 195-211.
2. J. K. Choi and L. J. Howe, "Histone Acetylation: Truth of Consequences?," Biochem.Cell Biol. 87, no. 1 (2009): 139-150.

Baf53A, BCoR, CBP, CBX5, CHD3, CHD4/Mi2 beta, CoAA, CtIP, FoxP3, HDAC1, HDAC2, HDAC3, HDAC6, HDAC7, KPNA2/RCH1, MBD3, MTA2, MYST1/MOF, PGC-1, Prohibitin, Rad9, RbBP4, RbBP7, SATB2, SDS3, SET, Sirt1, Sirt2, Sirt6, Sp1, TBLR1, Topo II Alpha, Topo II Beta.

Antibodies to Integrator Complex Proteins

2010-06-13T13:00:00.000-07:00

The integrator complex was identified in a study of DSS1 (deleted in split hand/split foot 1), a factor that associates with BRCA2 and the 19S proteasome (1). The core integrator complex is a 12 subunit complex (INTS1-INTS12) that associates with the C-terminal domain of the largest subunit of the RNA polymerase II (RNAPII) holoenzyme. In association with RNAPII, the integrator complex mediates the 3’ end processing of small nuclear RNAs U1 and U2. Recently, additional components of the integrator core complex have been identified (2). New members of the extended integrator complex include a phosphatase module (PPP1CB, PPP2CA/B, and PPP2R1A/B), four uncharacterized proteins (C12orf11, C15orf44, C7orf26, and C9orf80), two OB-fold proteins (OBFC2A and OBFC2B), and a set of zinc-finger proteins (ZMYND8, ZNF687, and ZNF589). The functions of some of the integrator subunits have been characterized and linked to a variety of cellular processes and signaling pathways. INTS11 is the catalytic subunit of the integrator core complex and is directly involved in the processing of U1 and U2 snRNA genes (1). INTS6/DICE1 has been implicated to play a role in Wnt signaling (3), and INTS3 has been shown to associate in a complex with human single-stranded binding protein (hSSB1) and possibly play a role in the DNA damage response independent of the integrator core complex (4).

References
1. D. Baillat et al., "Integrator, a Multiprotein Mediator of Small Nuclear RNA Processing, Associates With the C-Terminal Repeat of RNA Polymerase II," Cell. 123, no. 2 (2005): 265-276.
2. A. Malovannaya et al., "Streamlined Analysis Schema for High-Throughput Identification of Endogenous Protein Complexes," Proc.Natl.Acad.Sci U.S.A. 107, no. 6 (2010): 2431-2436.
3. S. Filleur et al., "INTS6/DICE1 Inhibits Growth of Human Androgen-Independent Prostate Cancer Cells by Altering the Cell Cycle Profile and Wnt Signaling," Cancer Cell Int. 9 (2009): 28.
4. J. R. Skaar et al., "INTS3 Controls the HSSB1-Mediated DNA Damage Response," J Cell Biol. 187, no. 1 (2009): 25-32.

INT1,
INT3 (FLJ21919)
INT4
INT5
INT6 (DDX26/DICE1)
INT7 (DKFZP434B168)
INT8 (FLJ20530/KAONASHI1)
INTS9 (RC74)
PP1CB
PPP2R1A
OBFC2A
SSB1
RNA Polymeriase II

Antibodies to Kinases in the MAPK Pathway

2010-06-07T12:38:00.000-07:00

The mitogen-activated protein kinase (MAPK) pathway can be activated by an assortment of extracellular stimuli such as growth factors, cytokines, and stress signals. Pathway activation leads to the coordination of a wide variety of cellular responses involving gene expression, proliferation, survival, apoptosis, metabolism, and differentiation. In mammals, there are five distinct groups of MAPKs: the extracellular signal-regulated kinases (ERKs), the c-Jun amino-terminal kinases (JNKs), the p38 isoforms, ERKs 3 and 4, and ERK5. In response to extracellular stimuli, ERKs, JNKs, and the p38 isoforms are activated by upstream MAPK kinases (MAPKKs) which are themselves activated by MAPKK kinases (MAPKKKs). For example ERK1 and ERK2 (ERK1/2) are activated by the upstream MAPKKs, MEK1 and MEK2 (MEK1/2). The MAPKKKs for MEK1/2 include A-Raf, B-Raf, Raf-1, and c-Mos which typically receive stimuli from growth factors via cell surface receptors. On the other hand, the p38 isoforms are activated by the MAPKKs, MEK3 and MEK6, which are mainly activated by environmental stress and inflammatory cytokine signals. The major substrates of MAPKs include MAPK-activated kinases (MKs) and transcription factors such as c-Jun, c-Myc, Max, CREB, p53, and ATF-2. The diverse biological functions influenced by MAPK signaling indicate that the MAPK network is complex. This complexity leaves volumes that require further study for a full understanding of the control of the MAPK modules as well as module cross-talk.

Selected Review
1. P. P. Roux and J. Blenis, "ERK and P38 MAPK-Activated Protein Kinases: a Family of Protein Kinases With Diverse Biological Functions," Microbiol.Mol Biol Rev. 68, no. 2 (2004): 320-344.

AKT1, AKT2, ERK1, ERK2, HPK1, JIK/TAOK3, MAP4K4/HGK, MEK1, MEK2, MEKK1, MEKK2, MST1,2/STK3,4, MST1/STK4, P38 MAPK, PAK1, PAK2, PKC-alpha, RAF1/c-RAF, RSK1, RSK2.

Antibodies to Proteins Involved in NF-Kappa B Signaling

2010-06-01T05:29:00.000-07:00

At the time of its discovery, NF-kappa B was hypothesized to represent a cell specific transcription factor required for B-cell immunoglobulin (Ig) gene expression; instead it was soon determined that NF-kappa B would encompass a much wider functional arena that involved the activation of a number of pleiotropic transcriptional programs.

Classically, NF-kappa B is described as a transcription factor that reprograms gene expression of the immune system in response to injury, infection, and stress. The NF-kappa B transcription factor exists as a dimer composed of pairs of monomeric subunits that are part of the Rel Family of proteins. The members that make up this family include NF-kappa B1/p50, NF-kappa B2/p52, RelA/p65, c-Rel, and RelB. To regulate its activity NF-kappa B associates with inhibitor proteins. These inhibitor proteins are members of the I kappa B family of proteins and include I kappa B-beta, I kappa B-delta/NF-kappa B2/p100, I kappa B-gamma/NF-kappa B/p105, I kappa B-epsilon, I kappa B-zeta, and Bcl3. Association with I kappa B proteins sequesters NF-kappa B in the cytoplasm to regulate its activity. The phosphorylation of I kappa B by the I kappa B kinase complex (IKK) results in the ubiquitination and degradation of I kappa B and the release and translocation of NF-kappa B to the nucleus where it can activate a vast number of genes.

Due to its combinatorial nature, its ubiquitous expression, and its ability to mediate pleiotropic responses, NF-kappa B has become one of the most studied transcription factors. Additionally, the dysregulation of NF-kappa B has been associated with a multitude of diseases and is being considered as a potential target of pharmacological intervention. These points illustrate that our understanding of NF-kappa B is far from complete and that it will continue as a subject of intense study.

c-Rel, CKII alpha, IKK-alpha, I kappa B-beta, IKK-beta, NF-kappaB1, NF-kappaB2, RelA/p65, RelB.

Antibodies to Proteins Involved in Notch Signaling

2010-05-26T08:15:00.000-07:00

Notch signaling plays a central role in cell fate determination during embryogenesis and adult tissue homeostasis (1 and 2). The mechanism of Notch signaling is highly conserved and consists of Notch receptors and transmembrane ligands which reside on neighboring cells and interact to achieve cell-to-cell communication. Signaling via the Notch pathway is iterative and functions in a variety of developmental events (2). The canonical Notch signaling pathway begins at the cell surface where an interaction between Notch and one of its ligands, such as Delta or Serrate on the surface of a neighboring cell, engages the Notch receptor to undergo a proteolytic event. Cleavage of the Notch receptor results in the release of the notch intracellular domain (NICD) from the membrane. The NICD translocates into the nucleus where it assembles into a transcriptional activation complex with RBP_J (also known as CSL or C-promoter binding factor) and Mastermind-like coactivator proteins for the regulation of gene expression. The critical role of notch signaling during development has been demonstrated in variety of model organisms such as flies, worms, and mice. In these organisms deficiencies in the Notch pathway result in embryonic lethality. In humans several diseases have been linked to defects in genes involved in notch signaling. Human T-cell acute lymphoblastic leukemia has been found to be caused by gain-of-function mutations in the Notch1 receptor leading to activation and oncogenic activity (3). The requirement of Notch signaling during development as well as the association between Notch signaling and human diseases establishes this pathway as a critical subject of study.

Selected Reviews
1. Wendy R. Gordon, Kelly L. Arnett, and Stephen C. Blacklow. The molecular logic of Notch signaling: a structural and biochemical perspective. J Cell Sci. (2008) October 1; 121(Pt 19): 3109–3119.
2. Ulla-Maj Fiúza and Alfonso Martinez Arias. Cell and molecular biology of Notch. Journal of Endocrinology (2007) 194:459-474.
3. Jon C. Aster , Warren S. Pear , Stephen C. Blacklow. Annu Rev Pathol. Notch signaling in leukemia. (2008) 3:587-613.

Notch1, Notch2, MAML1, MAML2, MAML3, NUMB, HDAC1, HDAC2, SMRT, CTBP1, CBP, p300.

Antibodies to Polycomb and Trithorax Group Proteins

2010-05-20T11:56:00.000-07:00

The development of body parts is determined by transcriptional activators encoded by homeotic (Hox) genes. From the stage of embryo to adult, the maintenance of Hox gene expression in either an active or silenced state is critical to normal development. The importance of these genes is indicated by the striking effects of their incorrect expression in flies. Incorrect expression causes homeotic transformations where one body part is transformed into the likeness of another. Polycomb group (PcG) and trithorax group (trxG) proteins are highly conserved key regulators of Hox genes. PcG and trxG proteins were originally discovered in Drosophila as mutant genes that resulted in homeotic transformations involving the thorax. PcG and trxG proteins work as opposing forces to maintain Hox gene expression by epigenetically modifying chromatin structure via polycomb/trithorax group response elements (PRE/TREs). PcG proteins maintain silenced states of gene expression, while trxG proteins maintain active states. Much of what is known about PcG and trxG function and their regulation of Hox gene expression has been established through the extensive study of the Drosophila model. Human homologs of PcG proteins and trxG proteins have been identified and found to be involved in similar processes controlling stem cell self-renewal and cell fate. A continuation and advancement of our understanding of the epigenetic regulation of gene transcription by PcG and trxG in humans will certainly lead to answers concerning how these factors contribute to stem-cell function and cancer formation.

Selected Reviews
1. L. Ringrose and R. Paro, "Polycomb/Trithorax Response Elements and Epigenetic Memory of Cell Identity," Development. 134, no. 2 (2007): 223-232.
2. V. K. Rajasekhar and M. Begemann, "Concise Review: Roles of Polycomb Group Proteins in Development and Disease: a Stem Cell Perspective," Stem Cells. 25, no. 10 (2007): 2498-2510.

ARID1A/BAF250, ASH1, ASH2, ASXL2, BMI1, BRG1/SMARCA4, CBX2, CBX7, CBX8, CHD4/Mi2beta, CHD7, EZH2, FALZ/BPTF, HDAC2, JARID1B, JARID1C, MLL1, MLLT1, p400, Pc2, PHC3, RbBP4, RbBP5, RbBP7, Sirt1, SMARCA1/SNF2L, SMARCA2/BRM, SMARCB1/SNF5, SMARCC2/BAF170, SUZ12, TRX2.

Antibodies to RRM-containing Proteins

2010-05-17T07:13:00.000-07:00

The RNA recognition motif (RRM) is a highly conserved RNA-binding domain and is present in one, two, or multiple copies in RNA-binding proteins. The RRM consists of a 90 amino acid domain that contains two consensus sequences termed ribonucleoprotein (RNP) domains. The first RNP is an eight amino acid sequence (RNP1) centrally located in the 90 amino acid RRM, while the second is a six amino acid sequence (RNP2) located at the N-terminus of the RRM. Proteins with an RRM motif are abundant and found mostly in eukaryotes. They are diverse in structure and function and are involved in all post-transcriptional processes such as pre-mRNA processing, splicing, RNA editing, mRNA export, translational regulation, and mRNA stability and degradation. In recent years, the RRM has been shown to also be involved in protein-protein interactions. Structural studies of RRM domains and the complexes they form with RNA and proteins have given some insight into the determinants of their binding specificity. Current information on RRM-containing proteins reveals that the role of the RRM in the cell may extend beyond a simple component of RNA recognition. Although a multitude of proteins bearing an RRM motif have been identified, the specific functions of many remain uncharacterized. Further biochemical and structural studies are needed to understand the evolution and functional diversity of RRM-containing proteins.

ACINUS/ACN1, Caper, CoAA, CPSF59, CPSF68, CSTF2T/TauCSTF64, CSTF64, DDX50, EIF3S4/eIF3G, EIF3S9/eIF3B, eIF4B, eIF4H, EWS, FUS, FUSIP1, G3BP1, G3BP2, GW182, hnRNP-H, hSET1, hSET1B, Matrin 3, NELFE, NONO, Nucleolin, PABP4, PGC-1, PGC-1beta, PSF/SFPQ, RBM5, RBM6, RBM8a, RBM9, RBM10, RBM12, RBM13, RBM15, RBM16, BM25/RED120, RBM26, RBM27, RBM34, RBMX2, SPF45, SAFB1, SAFB2, SART3/TIP110, SHARP, TAFII68, Tat-SF1, ZNF638/NP220.

Antibodies against Proteins in the Circadian Pathway

2010-05-13T14:54:00.000-07:00

On many levels circadian rhythm is central to animal physiology. Based on the daily 24-hour solar cycle and extending to axis-tilt and seasons, the circadian clock functions as an intrinsic time-keeper that coordinates the function of the major organs of the body. At the top of the hierarchy for circadian control lies the superchiasmatic nucleus (SCN) of the anterior hypothalamus. The SCN receives light signals via the retinohypothalamic tract (RHT) and functions as the master oscillator. As the master, the SCN synchronizes and coordinates slave oscillators located in other parts of the brain as well as in the peripheral organs. At the cellular level, the SCN is implicated to house distinct populations of neurons that hold the molecular machinery required for the function of the master clock. At the molecular level, the clock machinery consists of a group of transcription factors that function under positive and negative feedback loops and regulate gene expression via specific regulatory elements. Central to these feedback loops are the period (Per1, Per2, and Per3) and cryptochrome (Cry1 and Cry2) genes whose transcription is regulated by the CLOCK/BMAL1 heterodimeric transcription factor. Since this group of clock genes also has been found to be expressed with periodicity in tissues and organs peripheral to the SCN, there is a need to understand how the master and slave oscillators interact. Additionally, since the circadian rhythm has been linked to human health, it will be important to find specifically how these genes play a role in the development of disease.

BMAL1, CKI delta, CLOCK, CREB, Cry1, Cry2, DEC1, EAR2, EGFR, PHLPP, and Timeless.

Antibodies to Proteins in the SMC Complexes

2010-05-10T11:23:00.000-07:00

Structural maintenance of chromosomes (SMC) proteins are critical regulators of chromosome organization and function (1). This family of chromosomal ATPases is highly conserved from bacteria and archaeal species to vertebrates. There are at least six eukaryotic SMC proteins. The SMC proteins display a common primary architecture that consists of five structural domains. The five domains include two nucleotide-binding motifs termed the Walker A and Walker B motifs located at the N- and C-terminus, a central “hinge” domain, and two coiled-coil motifs that flank the central “hinge”. The SMC proteins associate in specific dimer pairs and arrange in an anti-parallel, two-armed structure that folds into a V-shape at the “hinge” domain. Three SMC complexes have been identified and are defined by the SMC subunits which associate to form a heterodimer and their respective associated proteins. SMC1 and SMC3 are part of the cohesin holocomplex which also includes at least two non-SMC subunits Scc1/mcd1/rad21 and scc3/SAs. The cohesin complex is required for holding together sister chromatids after replication in S-phase and throughout G2 phase. SMC2 and SMC4 are part of the condensin complex which also includes three non-SMC subunits: CAP-D2, CAP-G, and CAP-H. Condensin plays a critical role in mitotic chromosome condensation during prophase and metaphase and segregation of sister chromatids during anaphase. SMC5 and SMC6 are part of a not-yet-named octameric complex (2). The function of this complex is not fully understood; but it appears to be polyfunctional. The SMC5/SMC6 complex displays activities similar to cohesin and condensin, and it appears to be mainly involved in a variety of DNA repair pathways, replication, and recombination. The necessity of SMC proteins to the chromosome cycle has been well demonstrated, and more studies of these factors are needed for a better understanding of the complex dynamics of chromosome structure and function.

Selected Reviews
1. T. Hirano, "The ABCs of SMC Proteins: Two-Armed ATPases for Chromosome Condensation, Cohesion, and Repair," Genes Dev. 16, no. 4 (2002): 399-414.
2. G. De Piccoli, J. Torres-Rosell, and L. Aragon, "The Unnamed Complex: What Do We Know About Smc5-Smc6?," Chromosome.Res. 17, no. 2 (2009): 251-263.

SMC1, Phospho SMC1 (S957), Phospho SMC1 (S966), SMC2, SMC3, Phospho SMC3 (S383), Phospho SMC3 (S1083), SMC4, SMC5, SMC6, CAP-D2, CAP-D3, CAP-G, CAP-G2, CAP-H, CAP-H2.

Antibodies Against Ku70 and Ku80

2010-05-07T05:32:00.000-07:00

The Ku protein was originally identified as an antigen recognized by antibodies present in the serum of patients with autoimmune disease. The designation of this antigen as “Ku” is reported to be derived from the surname of a Japanese patient. The Ku antigen is made up of a heterodimer of two subunits of approximately 70 kDa and 80 kDa. Biochemical characterization of the Ku antigen, along with studies of the DNA-dependent serine/threonine protein kinase, DNA-PK, revealed that Ku protein associates with the ends of linear duplex DNA and is the regulatory component of DNA-PK (DNA protein kinase). Later it was established that Ku protein plays a central role in DNA double-strand break repair and transposition. In the presence of DSBs, Ku binds DNA ends and recruits the DNA-protein kinase catalytic subunit (DNA-PKcs) leading to phosphorylation events and further recruitment of DNA repair enzymes to the break site. Ku not only localizes to the nucleus as first described, but it is also found in the cytosol and on the cell surface. This discovery has broadened the probable functions of Ku antigen. In the cytosol, Ku plays a role in cell-survival; and on the cell surface, it is suspected to play a role in cell adhesion. The discovery of Ku as a cell surface protein has lead to the investigation of alternate roles of Ku in transformation and metastasis.

Selected Reviews
1. W. S. Dynan and S. Yoo, "Interaction of Ku Protein and DNA-Dependent Protein Kinase Catalytic Subunit With Nucleic Acids," Nucleic Acids Res. 26, no. 7 (1998): 1551-1559.
2. C. Muller et al., "The Double Life of the Ku Protein: Facing the DNA Breaks and the Extracellular Environment," Cell Cycle. 4, no. 3 (2005): 438-441.

Ku70
Ku80

Antibodies to Ubiquitin Specific Proteases

2010-05-05T08:39:00.000-07:00

Since the modest beginnings of its discovery in the late 1970’s, ubiquitination has emerged as a central and critical process involved in the regulation of virtually all cellular activities. Ubiquitin (Ub) is a small (76 kDa) highly conserved protein ubiquitously expressed in eukaryotic organisms. The conjugation of ubiquitin to proteins is an important means to regulate protein function by targeting tagged proteins for proteasomal degradation and modifying their activity. The modification of target proteins by ubiquitination is reversible, and the removal of Ub can rescue proteins from degradation or re-modulate their activity. The deconjugation of ubiquitin is accomplished by the deubiquitinating enzymes (DUBs). The majority of DUBs in the human genome belong to the ubiquitin specific protease (USP) subclass of DUBs. Structurally, USPs contain a common catalytic domain that consists of two short well-conserved motifs, called Cys and His boxes.

Similar to the action of ubiquitination being involved in most cellular activities, deubiquitination by USPs has also been established as a critical aspect of many cellular processes and is viewed as a critical regulatory mechanism in the cell. USPs have been noted to be involved in apoptosis, the cell cycle, cell division, DNA damage repair, chromatin and histone modification, cytoskeletal organization, neurodevelopment, RNA processing, transcription, translation, transport, signal transduction and the regulation of kinase activity. Research on the regulation of these enzymes is only in its beginning phase, and more information on this aspect of USP biology will help to further delineate the cycle of ubiquitination.

USP1, USP3, USP4, USP5/IsoT USP7, USP9X, USP10, USP11, USP14, USP15, USP16, USP19/ZMYND9, USP20/VDU2, USP24, USP28, USP32 USP33, USP34, USP35, USP36/DUB1, USP37, USP38, USP47, USP48.

Antibodies to the WD-repeat Family of Proteins

2010-05-03T07:14:00.000-07:00

The WD-repeat family of proteins is a large family of proteins whose members are structurally homologous but functionally diverse. The WD-repeat motif is defined by four or more repeating units of a conserved core of 40-60 amino acids that begins with a glycine-histidine (GH) dipeptide and ends with a tryptophan-aspartic acid (WD) dipeptide. Regions within the WD-repeat exhibit sequence heterogeneity and are likely responsible for imparting their functional diversity. WD-repeat-containing proteins perform in a spectrum of cellular activities which includes RNA processing, signal transduction, transcription, vesicular trafficking, cytoskeletal assembly, chromatin assembly, and various aspects of cell division.

The crystal structure of the beta subunit of the heterotrimeric G-protein has been resolved and has provided a structural characterization of the WD-repeat. Each of the seven repeats of the G-protein beta subunit folds into small anti-parallel beta-sheets that radiate from a central pseudo-symmetrical axis to form a beta propeller structure. The surfaces formed by the beta propeller structure are proposed to function as a scaffold capable of coordinating reversible protein-protein interactions.

Protein-protein interactions provide a source of complexity for the multitude of physiological processes that must be executed by eukaryotes for development and for the maintenance of homeostasis. Through their conserved beta propeller structure, WD-repeat-containing proteins contribute to this source of complexity and have thus established themselves as a critical subject of study.

APC4, BOP1, Bub3, CAF-1 p60, CDC20, COP1/RFWD2, Coronin 1, Coronin 2, CSTF50, DMWD, DTL/CDT2, EIF3S9/eIF3B, EML4, FBW7, GbetaL, Gemin5, GRWD1/WDR28, HERC1, LIS-1, LRWD1, NRIP, NUP214, PIP1/WDR84, PLRG1, PPP2R2B, PRP19/PSO4, RACK1, Raptor, RbBP4, RbBP5, RBBP7, RCD8, RFWD3, SEC31A, TBLR1, TRIM3/BERP, UTP18/WDR50, VprBP, WDHD1/AND-1, WDR3, WDR4, WDR5, WDR26, WDR33, WDR42A, WDR43, WDR44, WDR46, WDR62, WDR77/MEP50, WDR79, WDR91, ZFP106.

Antibodies against mTOR Pathway Proteins

2010-04-30T11:52:00.000-07:00

The mammalian target of rapamycin (mTOR) is a member of the ATM-related family of kinases. The mTOR kinase has been found to play a central role in integrating signals that relay information pertaining to the conditions of the extracellular environment. In response to energy stress, hypoxia stress, and the availability of nutrients, growth factors, and hormones, mTOR signaling directs the modulation of a number of cellular processes. The modulation of processes such as translation initiation, ribosome biogenesis, and transcription ultimately leads to responses in cytoskeletal dynamics and organization, cellular growth, proliferation, apoptosis, and autophagy. mTOR signaling is mediated by two mTOR complexes (mTORC): mTORC1 and mTORC2. The mTORC1 and mTORC2 are defined by the association of the mTOR kinase with either raptor (mTORC1) or rictor (mTORC2). Dysregulation of the mTOR pathway has been observed in a range of human malignancies; thus, the mTOR kinase is considered to be a promising target for cancer therapies.

Selected Reviews

1. K. Inoki et al., "Signaling by Target of Rapamycin Proteins in Cell Growth Control," Microbiol.Mol Biol Rev. 69, no. 1 (2005): 79-100.

2. D. A. Foster and A. Toschi, "Targeting MTOR with Rapamycin: One Dose Does Not Fit All," Cell Cycle. 8, no. 7 (2009): 1026-1029.

Bethyl Laboratories' Portfolio

4EBP1, AKT1, AKT2, AktS1/PRAS40, AMPK alpha 1, AMPK alpha 2, BRAF, eIF4B, eIF4E, ERK1, ERK2, GbetaL, HIF1-alpha, IRS2, MEK1, MEK2, mTOR, p70S6K, PDK1, PPP5C, PTEN, Raptor, RAF1/c-RAF, REDD1, Rictor, RPS6, S6K2, Sin1, Tip41, TSC1, TSC2.

Antibodies to SWI/SNF-like Chromatin-remodeling Protein Complexes

2010-04-29T07:54:00.000-07:00

The SMARCs (SWI/SNF-related, matrix-associated, actin-dependent regulators of chromatin), and BAFs (BRG1-associated factors), have been identified as components of the mammalian SWI/SNF-like chromatin-remodeling protein complexes. These multi-protein complexes are proposed to function as ATP-driven motors that translocate along DNA and destabilize nucleosomal structures to facilitate transcription factor binding. Two fractions of SWI/SNF-like complexes, SWI/SNF-A (BAF) and SWI/SNF-B (PBAF), have been isolated. The SWI/SNF-A (BAF) complex is defined by the presence of a BAF250/ARID1A subunit and either a SMARCA4/BRG1 or SMARCA2/BRM subunit, while the SWI/SNF-B (PBAF) complex lacks the BAF250/ARID1A subunit and contains a SMARCA4/Brg1 subunit and a polybromo/BAF180 subunit. In addition to these defining subunits, the SWI/SNF-A (BAF) and SWI/SNF-B (PBAF) complexes possess multiple subunits of other SMARCs and/or BAFs that are present in various combinations to provide the mammalian cell with a functional variety of SWI/SNF-like chromatin-remodeling complexes. This growing family of interacting proteins has also been shown to associate with the N-CoR (nuclear receptor co-repressor) deacetylase complex and various nuclear transcription factors. Current information indicates an important role for these complexes in the regulation of cell growth and proliferation. Future studies will likely demonstrate that the SWI/SNF proteins are important to many aspects of development and tumorigenesis.

ARID1B, BAF180/PB1, BAF250/ARID1A, BAF53A, HELLS, HMG20B/BRAF35, SMARCA1/SNF2L, SMARCA2/BRM, SMARCA3, SMARCA4/BRG1, SMARCAD1/ETL1, SMARCAL1, SMARCB1/SNF5, SMARCC1/BAF155, SMARCC2/BAF170, SMARCD1/BAF60a, SMARCE1/BAF57, SNF2h/ISWI.

Antibodies to Kinesins

2010-04-25T17:56:00.000-07:00

The kinesins (KIFs) make up a large superfamily of motor proteins that function in intracellular transport along microtubules. Kinesin activity has been linked to various cellular functions such as organelle and vesicle transport, mitotic spindle formation, chromosome segregation, and cytokinesis. Structurally, all kinesins contain a motor domain with microtubule and nucleotide binding sites that utilize ATP to target cargo along microtubule filaments. The kinesin motor proteins have been implicated in human disease. A mutation in the KIF1B locus is associated with Charcot-Marie-Tooth disease (CMT2A) which is characterized by a progressive dysfunction of peripheral neurons and a lack of synaptic vesicle transport in nerve endings and axons. Over forty members of the KIF family have been identified and well characterized. However, when one considers the multitude of cellular tasks involving the intracellular transport of cargo, many questions concerning KIF regulation remain unanswered.

KIF1B
KIF1C/LTXS1
KIF2A
KIF2C/MCAK
KIF4A
KIF10/CENP-E
KIF11/EG5
KIF13A
KIF14
KIF18A
KIF20A/RAB6KIFL
KIF22/KID
KIF23/MKLP1
KIFC1

New ELISA Kits released March

2010-04-22T18:00:00.000-07:00

Bovine IgG1 ELISA Kit

Dog IL-1 beta ELISA Kit

Horse IL-1 beta ELISA Kit

Human Alpha-1-Antitrypsin ELISA Kit

Human Hemoglobin ELISA Kit

New Polyclonal Antibodies Released in March

2010-04-22T17:27:00.001-07:00

New antibodies qualified for WB/IP: A20, BMAL1, CEE, CLOCK, Cortactin, Cry1, Cry2, Dcp2, DKC1, E4BP4, EYA4, FKBP3, FKBPL, FOXP1, Gamma-Tubulin, KIAA1688, Ku70, Ku80, LPP, MK5, MLK4, NuSAP, PIK3R2, RCC1, TFE3, TMEM192, WDR11.

New antibodies qualified for IHC: G9A/EHMT2, MKRN1, MKRN2, PPP1CA, PPP1CB, PPP1CC, PPP3CA, PPP5C, PRMT6, SFRS8/SWAP, USP36/DUB1, ZC3H13.

Antibodies to GTPase Regulators

2010-04-18T05:41:00.000-07:00

When a cell receives extracellular signals it may need to activate or turn on a number of downstream signaling pathways in order to initiate a particular cellular response. The downstream signals activated by growth factors, cytokines, hormones, cell-cell interactions, or physical stress can be complex and result in the modification of a number of cellular processes. These far reaching and complex effects can be achieved via a simple biochemical concept that involves proteins that function as pivotal molecular switches and interact with a multitude of effectors to turn on or turn off particular sets of responses.

The molecular switches are G-proteins, GTPases that hydrolyze GTP and cycle between two conformational states depending on whether GTP (guanine triphosphate) or GDP (guanine diphosphate) is bound. The GTP-bound GTPase is active, while the GDP-bound is inactive. In its active GTP-bound state, GTPases recognize and interact with target effectors that propagate downstream signals. Regulation of GTPase activity is achieved via activators called GEFs (guanine-nucleotide exchange factors) and inactivators called GAPs (GTPase-activating proteins). GEFs transduce signals from receptor tyrosine kinases, G-protein-coupled receptors, adhesion molecules, and second messengers and promote GTP binding to GTPases. GTP binding and signaling to target effectors is in turn terminated by the hydrolysis of GTP to GDP facilitated by GAPs. A third class of GTPase regulators, the GDIs (G-nucleotide dissociation inhibitors), also exists. GDIs are specific regulators of the Rho/Rac and Rab families of monomeric GTPases. GDIs influence G-protein signaling by binding the GDP or GTP form of the GTPase and preventing dissociation of the nucleotide.

It is apparent that the regulation of GTPase signaling is highly and carefully regulated. Hundreds of GTPase regulators have been identified and the G-protein signaling system is conserved well through evolution. The involvement of G-protein signaling in a wide variety of cellular activities has resulted in the study of GTPases and their regulators as therapeutic targets in a number of human diseases.

ArfGAP1, ArfGAP2, ArfGAP3, ARHGEF5, ARHGEF16, ASAP1, BIG1/ARFGEF1, BIG2/ARFGEF2. Borg2, BRAF, BCR, C3G, Cdc42GAP, CENTD1, CENTD2/ARAP1, CENTG3, CSN1, DAB2IP, Diaphanous 1, Diaphanous 2, DOCK4, DOCK9, DOCK10, DOCK11, DOCK180, ECT2, eEF2, EFTUD2/SNRP116, eIF5, eIF5B/IF2, EVI5, Filamin A, GAPex5, GBF1. GEF-H1. RLF1/p190RhoGAP, HERC1. HERC2. LARG., NF1, PARG1, PDZ-GEF1, PDZ-GEF2, PDZ-RhoGEF, PLEKHG2, Rab3-GAP1, RanBP1, RanBP2, RanBP3, RanBP8, RanGAP1. RASAL2, RCC1, Sin1, SIPA1, SOS1, SOS2, SRGAP1/ARHGAP13, TBC1D9/MDR1, TIAM1, TSC1, TSC2.

Antibodies to Subunits of the Cullin-E3 Ligases

2010-04-15T07:30:00.000-07:00

Ubiquitin-directed degradation by the 26S proteasome is central to the control and regulation of a multitude of cellular processes. The covalent attachment of polyubiquitin chains to target proteins is coordinated by three classes of enzymes: E1 ubiquitin-activating enzyme, E2 ubiquitin-conjugating enzyme, and E3 ubiquitin ligase. E1 is responsible for activating ubiquitin and passing it on to E2. E2 then assembles with E3 and transfers ubiquitin to a substrate protein. Multiple rounds of ubiquitin transfer occur to form polyubiquitin chains on the target protein and flag it for destruction. The E3 ligases play a critical role in this process because they determine the substrate specificity of the ubiquitination reaction. The two major types of E3 ligases are the cullin-RING-finger ligases (CRLs) and the anaphase-promoting complex/cyclosome (APC/C). The core complex of CRLs is composed of a cullin-homology domain-containing protein, a RING-H2 protein, and an adapter protein which facilitates interaction with substrate receptors. Seven cullin proteins have been identified in vertebrates: Cul1, Cul2, Cul3, Cul4a, Cul4b, Cul5, and Cul7. Studies indicate that each cullin protein associates with specific bridging adaptors and substrate receptors in the core complex to form substrate specific ligases. For example Cul1 associates with Skp1 and an F-box protein to form the SCF (Skp1, Cul1, F-box) ligase. In this complex, Skp1 bridges Cul1 to the F-box protein which functions as the substrate specific adaptor. Other Cul-based ligases include ECS (Elongin C, Cul2, SOCS), Cul4-DDB (DDB, Cul4, DCAF) and Cul3-BTB. A multitude of F-box, SOCS, BTB-domain, and DCAF factors have been identified and provide a critical means for assigning substrate specific activity to the E3 ligase function.

Selected Reviews

1. J. W. Harper, J. L. Burton, and M. J. Solomon, "The Anaphase-Promoting Complex: It's Not Just for Mitosis Any More," Genes Dev. 16, no. 17 (2002): 2179-2206.

2. J. Lee and P. Zhou, "DCAFs, the Missing Link of the CUL4-DDB1 Ubiquitin Ligase," Mol Cell. 26, no. 6 (2007): 775-780.

3. L. Pintard, A. Willems, and M. Peter, "Cullin-Based Ubiquitin Ligases: Cul3-BTB Complexes Join the Family," EMBO J. 23, no. 8 (2004): 1681-1687.

Bethyl Laboratories' Portfolio

AMBRA1, COP1/RFWD2, Cul3, Cul4a, Cul5, Cul7, DDB1, DTL/CDT2, FBW7, FBXO28, FBXO31, FBXO38, LIS-1, NRIP, PHIP, RBBP4/RbAp48, RbBP5, RbBP7, Senataxin, VprBP, WDR21A, WDR26, WDR28/GRWD1, WDR42A, WDR5, ZBTB40, ZBTB7/FBI-1, ZNF295.

Antibodies Against Proteins Associated with Microcephalic Disorders

2010-04-13T18:57:00.000-07:00

Brain size and cognitive ability sets humans apart from their closest evolutionary relatives. It is suggested that the phenomenon of encephalization in hominids may have involved gain-of-function changes in genes that have been associated with microcephalic disorders. Primary microcephaly, Seckel syndrome, and microcephalic osteodysplastic primordial dwarfism type II (MOPDII) are disorders that exhibit microcephaly, a human neurodevelopmental disorder characterized by reduced brain size. Microcephalic brains show loss of cellularity but near-to-normal architecture which strengthens the assertion that the associated genes may be responsible for the increase in human brain size through evolution. The genes associated with primary microcephaly include MCPH1, MCPH2, MCPH3/CDK5RAP2, MCPH4, MCPH5/ASPM, MCPH6/CENPJ, and MCPH7/SIL. Mutations in ATR and pericentrin have been shown to be the cause of Seckel syndrome and MOPDII, respectively. The products of some of the MCPH genes and the pericentrin gene have been localized to the centrosome and mitotic spindle. At the cellular level, these proteins play roles in the DNA damage response, the regulation of cell cycle progression, checkpoint regulation, and proliferation capacity. The exact role of these genes in neurogenesis is still not clear, and further studies of this group of proteins are required to assess their biological functions as well as provide insight into their role in hominid encephalization.

ATR

MCPH1

MCPH3/CDK5RAP2

MCPH5/ASPM

MCPH6/CENPJ

MCPH7/SIL

Pericentrin/Kendrin

Selected Reviews
1. G. K. Thornton and C. G. Woods, "Primary Microcephaly: Do All Roads Lead to Rome?," Trends Genet. 25, no. 11 (2009): 501-510.

2. B. Delaval and S. J. Doxsey, "Pericentrin in Cellular Function and Disease," J Cell Biol. 188, no. 2 (2010): 181-190.

Veterinary Cytokine ELISA kits

2010-03-09T11:53:00.000-08:00

Bovine CCL2 ELISA Kit

Cat IL-2 ELISA Kit

Horse IL-2 ELISA Kit

Pig CCL2 ELISA Kit

Pig IL-13 ELISA Kit

New Polyclonal Antibodies Released in February

2010-03-09T11:12:00.000-08:00

New antibodies qualified for WB/IP: ABCA2, ADRM1, AMBRA1, ARHGEF6, ARID3B, Beclin 1, BRD2, CNOT2, eIF2B5, HCA66, JMY, MCM7, NCBP2, Pumilio 1, Rubicon, SA1, SA2, TACC3, TOP1, Vps15,

New antibodies qualified for IHC: BCAS3, DC8, Gemin4, JARID1A/RBP2, KIF14, MAVS/VISA, MIS12, NCOA5/CIA, PAF49/CAST, PPP4R3 Beta.

New Cytokine Antibodies: Bovine CCL2, Bovine CXCL9, Bovine IL-2, Cat IL-2, Chicken IL-16, Dog IL-1 beta, Horse CCL2, Horse IL-2, Horse IL-4, Pig CCL2, Pig CXCL10, Pig IFN-beta, Pig IL-13.

Antibodies to Proteins in the Set1 Complexes

2010-02-03T06:34:00.000-08:00

Posttranslational modifications of histones are critical to the regulation of DNA-directed activities. Histone modifications include acetylation, sumoylation, phosphorylation, ubiquitination, and methylation (1). Histone lysine methylation serves as critical mark for the recruitment of specific factors for chromatin remodeling and the regulation of replication, repair, recombination, transcription, and RNA processing. The methylation of histone H3 occurs on five lysine residues (K4, K9, K27, K36 and K79) (1;2). H3K4 (histone H3, lysine 4)methylation is associated exclusively with actively transcribed genes and exists in a mono-, di- and tri-methylated form. The yeast COMPASS/Set1 complex was the first H3K4 methyltransferase complex identified and is composed of eight subunits. The yeast Set1 complex is strongly conserved in humans and shown to be nearly identical to the human Set1 complex (hSet1). In yeast, the COMPASS/Set1 complex is the only H3K4 methylase present; however in mammals, 6 homologs of Set1 have been identified: hSet1A, hSet1B, MLL1, MLL2, MLL3, and MLL4. These subunits exist in at least 4 distinct complexes: hSet1 complex, MLL complex, MLL2 complex, and MLL3/4 complex. Each complex contains distinct subunits as well as shared subunits (2;3). The yeast model has provided a wealth of information on the mechanism of H3K4 methylation, further studies of the homologous mammalian complexes will yield clues on the specific roles that histone modifications play in the regulation of gene expression.

Yeast COMPASS/Set1 Complex - Set1, Bre2, Swd1, Swd3, Sdc1, Swd2, Spp1.

hSet1 Complex - hSET1, ASH2, RbBP5, WDR5, hDPY-30, hSwd2, CXXC1, HCF1.

MLL Complex - MLL1 (TRX1), ASH2, RbBP5, WDR5, hDPY-30, HCF1/HCF2, Menin.

MLL2 Complex - MLL2 (TRX2) , ASH2, RbBP5, WDR5, hDPY-30, Menin.

MLL3/MLL4 Complex - MLL3/MLL4, ASH2, RbBP5, WDR5, hDPY-30, PTIP, PA1, ASC2, UTX.

Information adapted from Shilatifard (2008) 2 and Cho et al. (2007) 3

1. M. Lachner, R. J. O'Sullivan, and T. Jenuwein, "An Epigenetic Road Map for Histone Lysine Methylation," J Cell Sci. 116, no. Pt 11 (2003): 2117-2124.

2. A. Shilatifard, "Molecular Implementation and Physiological Roles for Histone H3 Lysine 4 (H3K4) Methylation," Curr.Opin.Cell Biol. 20, no. 3 (2008): 341-348.

3. Y. W. Cho et al., "PTIP Associates With MLL3- and MLL4-Containing Histone H3 Lysine 4 Methyltransferase Complex," J Biol Chem. 282, no. 28 (2007): 20395-20406.

New Cell Lysate Controls for Western Blot

2010-02-03T06:28:00.000-08:00

HeLa Whole Cell Lysate
CL300-100

HEK293T Whole Cell Lysate
CL300-101

NIH/3T3 Whole Cell Lysate
CL300-102

New Polyclonal Antibodies Released in January

2010-02-03T06:05:00.000-08:00

New antibodies qualified for WB/IP: APC7, BRCC36, CARMA1, CBX2, CBX7, CEP68, IBP160, MBD3, MCEF, PHF20, PRMT3, RACK1, REDQ5, RNF219, RuvBL2, SSA1, TFIP11, TMEM131L, Vinculin.

New antibodies qualified for IHC: BRG1/SMARCA4, DDX28, Histone H3, KARS, PIMT, PPP4R3, Rad52, SKIV2L2, TCF12, USP34.

New Agarose Immoblized antibodies: Actin, AKT2, Alpha Smooth Muscle Actin, Aurora A, BRG1/SMARCA4, CDK7, CRABP2, KLH, mTOR, Rictor, S6K2, Sin1, SMARCA2/BRM.

New Polyclonal Antibodies Released in December

2010-02-03T05:39:00.000-08:00

New antibodies qualified for WB/IP: ABCF3, ABI2, BRCC36, CCDC86, CDC37, CDC6, CDC7, CDK8, CEP152, EAF2, ELL2, MALT1, MERIT40, MIRK, NEDD4L, POGZ, SAP130, SF3A3, SF3B3, Sp3, SPATA2, SPF45, Spt16, WDR43.

New antibodies qualified for IHC: DJ-1, E2F1, eEF1G, eIF2alpha/EIF2S1, HIF-1 alpha, RAP80, RPL7, rpl7a/SURF3, TORC1, UBE1C.

New ELISA Kits

2010-01-13T10:29:00.000-08:00

Bovine IgA ELISA Kit
E11-121

Bovine IgM ELISA Kit
E11-101

Dog IgG ELISA Kit
E44-118

Mouse IgA ELISA Kit
E99-103

Mouse IgM ELISA Kit
E99-101

Pig IgA ELISA Kit
E101-102

Pig IgM ELISA Kit
E101-100

Pig IgG ELISA Kit
E101-104

Rabbit IgG ELISA Kit
E120-111

New ELISA Kits released in October

2009-11-08T13:37:00.000-08:00

Bovine CCL2 ELISA Kit
E11-800

Bovine Lactoferrin ELISA Kit
E11-126

Human Kappa ELISA Kit
E88-115

Human Lambda ELISA Kit
E88-116

Pig IL-13 ELISA Kit
E101-801

New Polyclonal Antibodies Released in October

2009-11-08T13:11:00.000-08:00

New antibodies qualified for WB/IP: ANKRD52, BAD, BHD, BORG4, BORG5, BRD3, CARS, CCDC28A, CENTD1, CRSP7, Cyclin Y, FBXO28, FBXO38, FOXC2, FOXO4, GMP synthase, HMGN1, MARCKS-like1, MAST4, MED13L, MED15, MEKK1, MTUS1, MYT1, NPM1, NUP133, p66alpha, Pc2, PLEKHG2, PPP1R14B, SPECC1, SRBC, STAT3, SUZ12, Stanniocalcin 2, TAF3, Tankyrase 1, UTX, ZNF639, ZNF768, ZNF8.

New antibodies qualified for IHC: Lasu1/Ureb1, RanBP1, RPC2, RPS6, SYK, ZBTB7/FBI-1.

New ELISA Kits

2009-10-10T16:00:00.000-07:00

Bovine IgG ELISA Kit
E11-118

Chicken IL-16 ELISA Kit
E33-800

Horse IL-2 ELISA Kit
E77-800

Mouse Albumin ELISA Kit
E99-134

Pig Albumin ELISA Kit
E101-110

Pig CCL2 ELISA Kit
E101-800

New Polyclonal Antibodies Released in September

2009-10-10T15:33:00.000-07:00

New antibodies qualified for WB/IP: AF4, ANKRD26, ARA70, ASPL, Ataxin-1, BRD7, BTBD12, BTF3, CapH2, CBX6, c-Cbl, CCDC82, CENP-T, CEP192, CEP76, Citron, DDX43, DHX57, DLG5, EB1, ECT2, Endophilin A2, FUSIP1, GW182, hSET1B, NUP188, p57kip2, p600, PGC-1beta, PHF23, Pig IL-13, PIN1, POLR3D, PRCC, PTP1B, RAD23B, RAI1, RBM34, Sec31A, Septin 9, SuFu, TFG, TIGAR, USP32, USP35, and YAP1.

New antibodies qualified for IHC: Bub1, DDX3, DDX6, FOXO3a, HDAC3, MST1,2/STK3,4, Paxillin, Rictor, SNX1, TPX2, and WSTF.

LOI and ART

2009-10-02T11:29:00.000-07:00

In a recent article published in Human Molecular Genetics, Katari et al. report that children conceived via assisted reproductive technology (ART) exhibit epigenetic differences in gene expression as a result of aberrant DNA methylation and gene imprinting (1). Developmental abnormalities as a result of embryo manipulation were first recognized in farm animals and described as ”large offspring syndrome” (2). The cause of this syndrome was later attributed to in vitro embryo manipulation and loss of imprinting (LOI) which involves the hypo- or hyper- methylation of genes and leads to aberrant repression or activation of the affected gene (in this case the IGF receptor). After the connection was made between ART, large offspring syndrome, and LOI in animals, it was hypothesized that human embryos conceived via ART might also experience imprinting defects. Studies investigating this possibility revealed that there was indeed an association between ART and disorders such as Beckwith-Wiedemann Syndrome and Angelman syndrome, two disorders previously known to involve LOI (3,4). The recent article by Katari et al. suggests that ART may have a global effect on DNA methylation that may be associated with the epidemiological observation that children conceived in vitro have a greater relative risk of low birth-weight, major and minor birth defects, and rare disorders involving imprinted genes.

Currently there are nine known human imprinting syndromes (5). Three have been associated with ART: Beckwith-Wiedemann, Angelman, and maternal hypomethylation syndrome. The remaining 6 syndromes include: Prader-Willi, Russell-Silver, maternal and paternal uniparental disomy of chromosome 14, pseudohypoparathyroidism type 1b, and transient neonatal diabetes. In addition to these syndromes, alterations in DNA methylation patterns have also been associated with cancer formation and neurobehavioral conditions. The association of epigenetic changes with human disease is most interesting because these changes do not involve genetic changes in the form of mutations in DNA sequences; but instead they involve changes in DNA structure that is inherited and stable through cell divisions. As the basis of genomic imprinting and epigenetic control of gene expression, understanding the process of DNA methylation and its heritable stability in the mammalian cell will be important to the study of a variety of human conditions.

Bethyl offers the following antibody products for your studies of DNA methylation:

ATRX, CTCF, DNMT1, DMAP1, HELLS, MBD2, MBD4, MCAF, and UHRF/NP95

References

1. Sunita Katari et al., "DNA Methylation and Gene Expression Differences in Children Conceived in Vitro or in Vivo," Human Molecular Genetics 18, no. 20 (2009): 3769-3778.
2. L. E. Young, K. D. Sinclair, and I. Wilmut, "Large Offspring Syndrome in Cattle and Sheep," Reviews of Reproduction 3, no. 3 (1998): 155-163.
3. M. R. DeBaun, E. L. Niemitz, and A. P. Feinberg, "Association of in Vitro Fertilization With Beckwith-Wiedemann Syndrome and Epigenetic Alterations of LIT1 and H19," Am.J.Hum.Genet. 72, no. 1 (2003): 156-160.
4. G. F. Cox et al., "Intracytoplasmic Sperm Injection May Increase the Risk of Imprinting Defects," Am.J.Hum.Genet. 71, no. 1 (2002): 162-164.
5. David J. Amor and Jane Halliday, "A Review of Known Imprinting Syndromes and Their Association With Assisted Reproduction Technologies," Human Reproduction 23, no. 12 (2008): 2826-2834.

Complete ELISA Kits for the Quantitation of Human Proteins

2009-09-22T11:12:00.000-07:00

Human ADAMTS13 ELISA Kit
E88-500

Human Albumin ELISA Kit
E88-129

Human Beta-2 Glycoprotein 1
E88-142

Human IgA ELISA Kit
E88-102

Human IgE ELISA Kit
E88-108

Human IgG ELISA Kit
E88-104

Human IgM ELISA Kit
E88-100

Human Lactoferrin ELISA Kit
E88-143

Human Transferrin ELISA Kit
E88-128

Kiss of Death or Gift of Life?

2009-09-11T06:55:00.000-07:00

It used to be that a protein that was ubiquitinated was a protein receiving the kiss of death. It determined that it would be marked for destruction and then sent to a proteasome to die. But as bad as it may sound, this is actually necessary and all good…because from death comes life (insert angels singing here). Ubiquitination and the regulated destruction of proteins are central to the life of a cell and to almost every aspect of a cell’s activities. And recently, it has been discovered that a protein that is sentenced to destruction by a kiss of death might not only maintain the life of a cell, but may actually be spared and gifted to receive life itself. The interest in ubiquitin and ubiquitination has recently been turned to its proteasome-independent roles and to the observation that ubiquitin can also function as a direct activator of signal transduction proteins (1;2). The most recent examples come from two manuscripts published very recently in Science and Nature.

In the August 28 issue of Science, Yang et al. from the UT M.D. Anderson Cancer Center in Houston, showed that in response to growth-factor stimulation, the ubiquitination of AKT on lysine-63 occurs and results in its phosphorylation, membrane recruitment, and activation of AKT (3). In these studies, TRAF6, an E3 ligase already implicated in the activation of signaling pathways, was determined to be the direct E3 ligase mediating Akt ubiquitination and activation.

The second manuscript demonstrating the involvement of ubiquitin in the activation of kinases was also reported by a Texas group (Yeee-Ha!). In this September 3, 2009 Science publication, Xia et al. from UT Southwestern, not only showed that ubiquitin was involved in the activation of the kinase TAK1 (transforming growth factor beta-activated kinase 1, also known as MAP3K7), but that free unanchored Lys 63 polyubiquitin chains catalyzed by TRAF6 and not the ubiquitination of the TAK1 complex itself were responsible for the direct activation of TAK1 (4). They further demonstrated and proposed a mechanism for the activation of TAK1 by free polyubiquitin chains. In this model, binding of unanchored polyubiquitin chains leads to the oligomerization of the TAB2 (TAK1 binding protein 2) or TAB3 subunits of the TAK1 complex and the subsequent autophosphorylation and activation of TAK1. This report is most interesting as it presents a novel mechanism for the activation and regulation of protein kinase activity. Additionally, it presents yet another level of the already complex ubiquitination system and the regulation of proteins by post-translation modifications.

Bethyl has available products for your studies of TAK1, AKT, TAB3, IKK-alpha, IKK-beta, and IsoT

Additionally, Bethyl’s portfolio of antibodies against proteins involved in the ubiquitin cycle include:
ANKIB1, APC4, APC6/CDC16, Apollon/BIRC6, APC1, APC5, APC8/CDC23, BAP1, BARD1, BBAP/DTX3L, BRCA1, BRE, CBX8, CDCA3/TOME1, CDT2/DTL, CHIP/STUB1, COP1/RFWD2, CSN4, CSN5, CSN7b, Cul3, Cul4a, Cul5, Cul7, DDB1, DUB1/USP26, DUB1/USP36, E4F1, E6AP, EDD1, FBW7, FBXO11, FLRF/RNF41, GCN2, H7-AP1/PARC, HERC1, HERC2, HERC4, HRS, Iso7/UPS5, Itch, JHDM1A, Lasu1/Ureb1, MIB2/Skeletrophin, MKRN1, MKRN2, NICE4/UBAP2L, NP95/UHRF1, PAM, PHD1, PHD2, PHD3, Pirh2, Plant Ubiquitin, PRP19/PSO4, Rad18, Rad6, RNF20, RNF40, Roquin, Senataxin, Sirt2, SUG1/TRIP1, TBLR1, TIF1gamma/TRIM33, Topo II Alpha, TRIM37, TRIP12, TSC1, UBCH7, UBE1, UBE1C, UBE2O, UBE2T, UBE4B, UFD1, USP1, USP10, USP11, USP14, USP15, USP16, USP19/ZMYND9, USP20/VDU2, USP24, USP28, USP3, USP33, USP34, USP37, USP38, USP4, USP47, USP48, USP7, USP9X, VCP, VprBP, and WRNIP1.

Literature Cited

1. D. Mukhopadhyay and H. Riezman, "Proteasome-Independent Functions of Ubiquitin in Endocytosis and Signaling," Science. 315, no. 5809 (2007): 201-205.
2. Z. J. Chen and L. J. Sun, "Nonproteolytic Functions of Ubiquitin in Cell Signaling," Mol.Cell. 33, no. 3 (2009): 275-286.
3. W. L. Yang et al., "The E3 Ligase TRAF6 Regulates Akt Ubiquitination and Activation," Science. 325, no. 5944 (2009): 1134-1138.
4. Z. P. Xia et al., "Direct Activation of Protein Kinases by Unanchored Polyubiquitin Chains," Nature. 461, no. 7260 (2009): 114-119.

New Polyclonal Antibodies Released in August

2009-09-04T10:47:00.000-07:00

New antibodies qualified for WB/IP: ABLIM1, AKT2, ARID2, ARID4A, ARID4B, ARID5B, BAP1, CAMSAP1, CEP135, CKAP4, DAP5, DDX27, DHX30, DOCK4, FAD24, GNL1, Haspin, HBO, MID1, MLLT1, Nestin, OBFC2A, POP1, RAD51AP1, RBM5, RBMX2, SAP97, SASH1, Sepharase, SET, SMG9, TAB3, TFIIS, WDR26.

New antibodies qualified for IHC: ABCF2, EphA2, MTA3, RAP1, Tat-SF1.

New Antibodies for Veterinary Research

2009-08-31T05:45:00.000-07:00

Antibodies qualified for Western Blot and ELISA:

Bovine CCL2, Chicken IL-16, Horse IL-2, Horse IL-4, Pig CCL2, Pig IL-13

New Polyclonal Antibodies Released in July

2009-08-13T05:29:00.000-07:00

Antibodies qualified for WB/IP: Abraxas, AF17, AF6, ARHGEF5, Axl, BCCIP, CENTG3, Cul5, FANCD2, JAK2, MARK3, MAST2, MASTL, MINK1, MKL1, NOPP140, PBK, REDD1, RelB, SMG7, SPHK1, TOPORS, and TRX2.

Antibodies qualified for IHC: Artemis, CC2D1A, EMSY, INT7, INT8, KIAA0082, MAD2, Phospho RPA32 (S33), Phospho RPA32 (S4/S8), RFC3, RPA32, RPA70, Tyro3, Ubiquitin, WRN, and ZEB1.

Self-Renewal

2009-07-08T08:48:00.000-07:00

No bars on your cell phone, no connection for mail, just warm sand on your soles, a salty breeze on your skin, and a frosty coconut concoction cooling you from the inside out. Then the sun sets west pulling down its heat, leaving only cool stars on the ocean that break apart at the shore. And under the tone of fizz and breaking waves, Mr. Marley is in the distance, telling you in a most lovely and melodic manner…”turn your lights down low”. The day is done and all that will be left is the relaxing energy that will become your self-renewal.

Now queue the sound of a turntable needle ripping across a vinyl record and then drop-kick yourself into reality (Ouch!): office phone ringing, inbox bulging, the rats closing in. How grand it would be to have some of that warm sand under our feet and a more than frequent chance at self-renewal. To be able to rejuvinate ourselves at our most basic cellular presence and leave us with no worries. But as the ripping sound continues, self-renewal at that cellular level actually proceeds; but it won’t transport us to that place far away. Instead it may, just as importantly, re-populate our tissues and lead us to clues about regenerative medicine and promising cancer therapies.

The concept of self-renewal (in the geeky sense) applies to adult pluripotent stem cells as well as embryonic stem cells. Self-renewal allows a cell to maintain itself in an undifferentiated state until it receives signals to commit to a particular lineage. I might think that the people in the lab of Steve Elledge probably haven’t had a dose of that grand level of self-renewal in a long time (they appear to not be allowed to sleep… how else is it possible to publish so prolifically!). But their hard work as well as that of others has paid off and contributed to our progress of getting to the bottom of how stem cells remain pluripotent and provide a constant source of cellular self-renewal. Hu et al., of the Elledge lab at Harvard Medical School, set out to identify genes essential for self-renewal in mouse embryonic stem (ES) cells. The group approached this by carrying out a genome-wide RNAi screen in mouse cells using a functional assay that employed a very clever GFP reporter system in which GFP expression correlated with ES cell identity. This screen led to the re-identification of known and suspected ES cell self-renewal genes as well as the identification of others not yet implicated in the network. Of the over 100 newly identified factors they selected two transcriptional regulators for further investigation since knockdown led to relatively strong phenotypes. Cnot3 and Trim28/KAP-1 were the two factors investigated and found to regulate self-renewal via the modification of chromatin structure. Further studies of human Cnot3, Trim28/KAP-1 and their associated proteins will likely lead to the identification of new stem cell pathways. And perhaps when we figure it all out, we can just relax in an ocean breeze.

Bethyl has rabbit polyclonal antibodies against human Cnot3 and Trim28/KAP-1 available for your studies. Antibodies are also available to the self-renewal proteins nanog, SOX2, RIF1, HP1-beta, and REST.

Oh Shi-gella!

2009-06-22T13:55:00.000-07:00

“Oh, Shigella”, words you don’t want to say when you are on a trip out of the country. Pathogenic bacteria such as Shigella and E. coli found in water or food contaminated by feces are one cause of the dreaded code brown emergency. Shigella is a gram-negative rod-bacterium related to E. coli and Salmonella. Shigella, as well as many other pathogenic bacteria, colonizes intestinal epithelium causing cell death and irritation which results in an inability of the epithelium to absorb water; so what can’t come in must go out. Shigella has been shown to deliver factors to epithelial cells that facilitate their infection of the intestinal epithelium. A study published last month in Nature (1)reveals that one such bacterial factor, OspE, inhibits sloughing of the intestinal cells allowing more efficient infection by the bacteria. OspE is able to promote host epithelial cell adherence to the basement membrane by interacting with the cellular protein, integrin-linked kinase (ILK). ILK plays a critical role linking the extracellular matrix to the actin cytoskeleton and signaling complexes. This study shows that OspE promotes retention of ILK at the membrane and facilitates and stabilizes focal adhesion formation. Stabilized focal adhesions inhibit normal rates of epithelial cell exfoliation, which functions as an important barrier to infection, allowing the bacteria to acquire and maintain an infectious foothold. The authors relate that the discovery of small molecules that might block ILK and OspE interaction may provide effective therapies for bacterial infections that use similar mechanisms.

Bethyl Laboratories has antibodies available against human ILK for use in WB, IP and IHC.

Reference List
1. Kim, M., M. Ogawa, Y. Fujita, Y. Yoshikawa, T. Nagai, T. Koyama, S. Nagai, A. Lange, R. Fassler, and C. Sasakawa. 2009. Bacteria hijack integrin-linked kinase to stabilize focal adhesions and block cell detachment. Nature. 459:578-582.

New Polyclonal Antibodies Released in May

2009-06-05T11:15:00.000-07:00

Antibodies qualified for WB/IP: AKT1, ARIP4, AKT1, ASF, ASAP1, BCR, C1orf55, CCDC6, CLASP1, Cyclin L1, ERK1, ERK2, FANCB, FBXO31, GAPex5, GIT1, GIT2, GSK3-beta, hPrp3p, INT3, LAP1B, LARP1, NF-YA, NF-YC, Notch2, PAR4, PHIP, Phospho RelA (S529), Phospho RelA (S536), Phospho RelA (S468), PRKCBP1, QSER1, RASAL2, SAM68, SAS10, SMC3, Rab3-GPA1, RALY, RASAL2, SAS10, SRcyp, THOC5, U2AF35, VPS4B, and Ymer.

Antibodies qualified for IHC: ABCB9, BRD4, Ki-67, and TCEB3.

New Business for BMI1

2009-05-29T14:26:00.001-07:00

Last week in Nature, a collaborative study from the NIH revealed an unexpected role for the polycomb complex repressor protein, BMI1 (B lymphoma Mo-MLV insertion region 1 homolog). BMI1 is a core component of the polycomb group (PcG) multiprotein PRC1 complex that functions as a transcriptional repressor of gene expression during development. The BMI1 gene was first isolated as an oncogene that cooperated with c-Myc to cause lymphomas in mice, and it has been found to be widely expressed in diverse types of human cancers. Studies of BMI1 deficient mice have shown that it is essential for the self-renewal and maintenance of hematopoietic and neural stem cells. Previous studies of BMI1 have concluded that the effects of BMI1 deficiency might be mediated by the de-repression of the Ink4a/Arf (Cdkn2a) locus which encodes two regulators of the cell cycle, p16INK4a and p14ARF. This most recent study indicates that there are additional pathways regulated by BMI1 that may be responsible for the BMI1 null phenotype. The authors demonstrate that BMI1 is involved in the regulation of reactive oxygen species (ROS) levels. Cells from Bmi1-/- mice exhibited increased levels of ROS and impaired mitochondrial function. Additionally, the administration of antioxidants was able to rescue thymocyte function as well as the “failure-to-thrive” phenotype in Bmi1 -/- animals. The study concludes that de-regulation of mitochondrial function and activation of the DNA damage response pathway involving Chk2 is partially responsible for the phenotype of Bmi1-/- mice and that the role of BMI1 in stem cell biology is related to its regulation of mitochondrial function.

Bethyl Laboratories’ catalog of polyclonal antibodies includes products for your studies of human BMI1, p16INK4a, p14ARF, and Chk2.

New Polyclonal Antibodies Released in April

2009-05-11T19:42:00.000-07:00

Antibodies qualified for IP/Western Blot: ABC50, ABCB9, Alpha Smooth Muscle Actin, ArfGAP1, ArfGAP2, ARG, ASXL2, BAT3, Beta-catenin, BRD4, CKI alpha, Crk-II, Cyclin C, DRIM, eIF3A/eIF3S10, EphA2, G3BP2, G3BP2, Itch, KSRP, Myosin-10, NOL1, OXR1, PACT, Phospho ZAP70-Y319, Phospho ZAP70-Y493, Phospho ZAP70-Y493/Y494, PHRF1, PMP70, Prohibitin, RanGAP1, Shc1, SIPA1, Tat-SF1, TIEG1, UVRAG, ZAK.

Antibodies qualified for IHC: DHX15, DRIL1, Drosha, E1B-AP5, ERF, Phospho EZH2 (S21), Phospho RNA Polymerase II (S5), USP1.

Drosha Takes Center (Microscope) Stage

2009-05-11T19:17:00.000-07:00

Detection of Human Drosha by IHC.
Sample: FFPE section of human ovarian tumor. Antibody: Rabbit anti-Drosha (Cat. No. IHC-00389) used at a dilution of 1:250. Detection: DAB staining using IHC Accessory Kit (Cat. No. IHC-101). Epitope Retrieval Buffer-High pH (IHC-101J) was substituted for Epitope Retrieval Buffer-Reduced pH.

Earlier this year in February, Bethyl Laboratories introduced an anti-Drosha polyclonal antibody (pAb) validated for use in western and IP assays. This month anti-Drosha makes another appearance, this time on the microscope stage as part of Bethyl’s line of immunohistochemistry (IHC) products. Bethyl’s newest anti-drosha (IHC-00389) pAb product has been validated and qualified for the detection of Drosha in human and mouse FFPE samples. All of the antibodies in Bethyl’s line of IHC products have been formulated and optimized for use on FFPE samples under standardized conditions using the reagents provided in Bethyl’s easy to use and convenient IHC Accessory Kit. The Bethyl IHC Accessory Kit is designed for immunostaining FFPE sections with primary antibodies made in rabbit. The kit utilizes an anti-rabbit secondary IHC antibody that eliminates the streptavidin- or avidin-HRP step as well as the blocking step for endogenous biotin that is required when using biotinylated secondary antibodies. In addition to its reliability and sensitivity, the Bethyl IHC accessory kit is also extremely economical: for about $1.26 per slide, one kit will stain at least 250 slides. For more technical information on the IHC kit please see the product data sheet.

For the detection of the Drosha target with anti-Drosha IHC pAb, epitope exposure with a Tris-EDTA pH 9.0 buffer is recommended. A reduced pH epitope retrieval buffer is included in the IHC Accessory Kit, however the high pH buffer can be purchased separately (Epitope Retrieval Buffer Concentrate- High pH IHC-101J).

Antibodies to ATM and ATR Signaling Proteins

2009-04-21T06:42:00.000-07:00

Exposure to mutagenic agents results in DNA damage that must be rigorously dealt with in order to prevent the cell from propagating mutations that can result in genomic instability and malignant transformation. To address the constant threat of genotoxic stress, animal cells have evolved a DNA damage response that is governed by a complex and highly-branched signaling network. The components of this network function to sense DNA lesions and transduce cellular signals that activate transcriptional programs for the initiation of DNA repair, cell death, or cell cycle arrest.

There are three cell cycle checkpoint pathways that arrest or delay DNA synthesis when genetic damage is sensed. The G1, S, and G2 cell cycle checkpoints are primarily regulated by the ATM (ataxia telangiectasia, mutated) and ATR (ATM and Rad3-related) protein kinases. ATM and ATR are members of the PI-3 family of serine-threonine kinases and function as essential links between the sensors and effectors of the DNA damage response. The roles of ATM and ATR partially overlap and are cooperative; however they are also known to play distinct roles in protecting the cell from DNA damage. ATM is mostly responsible for sending signals from DSBs (double-strand breaks) induced by ionizing radiation while the closely related ATR responds to UV damage or stalled replication forks.

ATM and ATR are known to phosphorylate common as well as specific substrates to activate checkpoint signaling. The list of reported substrates of these kinases is sizable and continues to grow. The ever-expanding and elaborate downstream network of ATM and ATR signaling illustrates both the versatility and complexity of this pathway and presents many directions of research that will require further exploration for a complete understanding of the DNA damage response. Even more paths of research remain for understanding the upstream events that take place at the level of sensing DNA damage and grasping how this is linked to ATM and ATR activation.

Bethyl Laboratories' current portfolio of antibodies to ATM and ATR Signaling Proteins:
4EBP1, 53BP1, phospho 53BP1 (S25), Artemis, ATF2, ATM, ATR, ATRIP, BID, phospho BID (S61), phospho BID (S78), BRCA1, phospho BRCA1 (S1189), phospho BRCA1 (S1280), phospho BRCA1 (S1387), phospho BRCA1 (S1423), phospho BRCA1 (S1457), phospho BRCA1 (S1466), phospho BRCA1 (S1524), cAbl, CDC25C, Chk1, phospho Chk1 (S317), Chk2, CtIP, E2F1, FUS, gamma-H2AX, H2AX, HdmX/MDM4, phospho HdmX/MDM4 (S403), MCM3, MDC1, Mre11, NBS1, p53, PP2A, PPP5C, Rad17, phospho RAD 17 (S645), Rad50, RPA32, phospho RPA32 (S4/S8), phospho RPA32 (S33), SMC1, phospho SMC1 (S957), phospho SMC1 (S966), Sp1, TRF2, WRN.

got lactoferrin?

2009-04-13T05:33:00.000-07:00

Got Milk? It’s a catchy slogan designed to sell more dairy products and distract drivers with attractive milk-moustache wearing models on bill boards. However, the more important question may be; Got Lactoferrin? Lactoferrin is found in high concentrations in many exocrine secretions. It is the second most abundant protein in human milk, and the secondary granules of neutrophils are loaded with the stuff. Lactoferrin appears to be important in myriad biological processes, including cell growth and development, regulation of immune response and iron absorption. Its’ immunomodulatory role may prove to be its’ most salient property. (1)

Wolf et al. report on the immunododulatory properties of lactoferrin in their article entitled , oral lactoferrin results in T Cell-Dependent tumor inhibition of head and neck squamous cell carcinoma in vivo. (2) In-vitro data suggests that LF induces direct cell cycle arrest, possibly via suppression of proinflammatory cytokines and NF-kB. (3) However, this article suggests that the anti-tumor effect is the result of immunomodulatory properties and the resultant lymphocytic infiltration of the tumors.

The potential antimicrobial effects of lactoferrin were lauded in a 2008 article published in the Journal of Medical Microbiology, where lactoferrin demonstrated synergy with vancomycin and nafcillin against coag-neg staph and with ampho B and fluconazole against Candida albicans.(4)

In an article by de la Rosa et al., in the Journal of Immunology, recombinant human lactoferrin resulted in increased maturation of monocyte-derived dendritic cells in culture.(5) Further supporting the role of lactoferrin as key in modulating the immune response .

Lactoferrin is emerging as a promising therapeutic adjunct in the treatment of infectious, inflammatory and malignant disease. Again, adding evidence that mother’s milk is “nature’s perfect food”.

Bethyl Laboratories is pleased to announce the release of our Human Lactoferrin ELISA Kit. The kit comes complete with sufficient components to quantitate Human Lactoferrin protein concentration in up to 40 samples, tested in duplicate.

Reference List:
1. Orla M. Conneely Antiinflammatory Activities of Lactoferrin J. Am. Coll. Nutr. 2001; 20 (90005) : 389S-395S.
2. Jeffrey S. Wolf, Guoyan Li, Atul Varadhachary, Karel Petrak, Mark Schneyer, Daqing Li, Julina Ongkasuwan, Xiaoyu Zhang, Rodney J. Taylor, Scott E. Strome, and Bert W. O'Malley, Jr. Oral Lactoferrin Results in T Cell–Dependent Tumor Inhibition of Head and Neck Squamous Cell Carcinoma In vivo Clin. Cancer Res. 2007; 13 (5) : 1601-1610.
3. Jun-Ichi Togawa, Hajime Nagase, Katsuaki Tanaka, Masahiko Inamori, Tadashi Umezawa, Atsushi Nakajima, Makoto Naito, Shinobu Sato, Toshifumi Saito, and Hisahiko Sekihara Lactoferrin reduces colitis in rats via modulation of the immune system and correction of cytokine imbalance Am J Physiol Gastrointest Liver Physiol 2002; 283 (1) : G187-G195.
4. Mohan Pammi Venkatesh and Liang Rong Human recombinant lactoferrin acts synergistically with antimicrobials commonly used in neonatal practice against coagulase-negative staphylococci and Candida albicans causing neonatal sepsis J. Med. Microbiol. 2008; 57 (9) : 1113-1121.Gonzalo de la Rosa, De Yang, Poonam Tewary, Atul Varadhachary, and Joost J. OppenheimLactoferrin Acts as an Alarmin to Promote the Recruitment and Activation of APCs and Antigen-

Antibody to Mouse Ki-67

2009-04-05T17:09:00.000-07:00

Antibodies against the Ki-67 antigen have become reagents with widespread applications in diagnosis, prognosis, research, and drug discovery. Anti-Ki-67 antibodies recognize a biomarker specific for proliferating cells, and their use in many aspects of the study of cell biology is considered invaluable.

Bethyl Laboratories is excited to announce the release of a new anti-murine-specific Ki-67 rabbit polyclonal antibody. This new anti-Ki-67 product has been validated and qualified for use in immunohistochemistry in formalin-fixed paraffin-embedded mouse specimens. When considering price and quality, investigators now have an alternate source to consider for their anti-K-i67 antibody needs.

Mouse models are vital to understanding the cellular basis of human disease. They have been especially valuable to the study of human cancer and the screening of anti-cancer drugs. Since cancer is a disease characterized by malignant growth and proliferation of cells, the study of cancer and potential cancer therapies often involves the measurement of proliferation in model neoplasms. The determination of the growth fraction in neoplasms allows pathologists to correlate proliferation indices with histologic grade and prognosis, and to monitor the progress of experimental targeted therapies. Several methods for determining the proliferation index of tumors can be used such as enumerating mitotic figures via light microscopy, assaying DNA synthesis by incorporation of DNA precursors or nucleotide analogs (e.g. tritiated thymidine or bromodeoxyuridine), measuring DNA content by flow cytometry, and immunohistologically detecting the expression of biomarkers for proliferation. Due to its ease and precise reflection of proliferation, anti-Ki-67 immunohistochemistry is acknowledged as a preferred method for the measurement of proliferation indices.

In addition to furthering the progress of the study of cancer models in mice, mouse-specific Ki-67 antibodies are popular to those who study tissue regeneration and nerve tissue development and growth. The introduction of this new product will provide another quality reagent from Bethyl Laboratories that will advance our knowledge of the basic biology of disease and disease therapy.

New Polyclonal Antibodies Released in March

2009-04-05T16:52:00.000-07:00

Antibodies qualified for IP/Western Blot: ARHGEF16 C3G CDC7 CEP97 CENP-E CEP97 ch-TOG CKII beta CLP36 Cyclin K Dicer eIF2A FANCA GBF1 IKK-alpha IQGAP1 LARG LMP1 NDE1 PA1 PACSIN3 PDZ-GEF1 PDZ-RhoGEF Pol kappa SSB1 vPARP ZBTB40

Antibodies qualified for IHC: CDK2 COBRA1 Ki-67 NELFA NELFE p66beta/GATAD2B Paf1 Parafibromin PARP10 Pds5B PHLPP Phospho RNA Polymerase II (S2) Rad9

Just Tell It Like It Is

2009-03-30T10:18:00.000-07:00

Okay it’s been a month since daylight savings time took effect and I think that just now my body is starting to get on schedule. My son is also finally getting used to the change, but at the beginning he was so confused with the concept we had multiple conversations concerning the topic. Here’s one of them:

He says: “Mom, why is it earlier but later”?
My answer: “’Cause it’s later but earlier”. (huh?)
He says: “But what time is it really”?
I tell him: “It’s whatever time the clock “really” says it is”.

He keeps on (because this is the job of 7 yr olds, to keep asking the same question in hopes of hearing a different not-so-smart-aleky answer):

“But why does it LOOK like I should still be playing outside, but YOU’RE telling me it’s time to bathe and get to bed”?
I say: “because maybe I’m trying to get you out of my hair”.

One day he’ll learn to both not keep asking his mother the same question in hopes of getting the answer he wants, and exactly why it is so hard to get used to the whole daylight savings thing. But perhaps I should have just explained scientifically to him what is going on and why it seems “earlier but later” –HA! Are you kidding me? What a mess that would be. As it is I still have to explain the concept of time by relating it to the number of SpongeBob episodes that fit into an hour.

When we change the clocks ahead one hour in the spring it is a fact that it is not easy for the human body to physiologically adjust to the change, and the difference of only one hour truly has a great impact on our internal hard-wired biological clock or circadian rhythm. Many metabolic and physiological processes display circadian oscillations. Some scientists feel that forcing changes to our natural circadian rhythm could have lasting effects on our health and that disruption of these rhythms may be linked to obesity, diabetes, insomnia, depression, cardiovascular disease, and cancer (no wonder I feel like hell). In the March 12, 2009 issue of Science magazine, a group from UC Irvine has shown further evidence that our metabolism is closely tuned with our circadian rhythm. In this study Nakahata et al. (1) show how the proteins CLOCK, SIRT1, and NAMPT/visfatin function together as part of the circadian clock. CLOCK functions as an acetyltransferase for histones and promotes circadian chromatin remodeling. SIRT1 functions as a deacetylase that counteracts the histone acetlytransferase activity of CLOCK. SIRT1 activity has been shown to be dependent on intracellular NAD+ levels. This led the group to investigate whether NAD+ levels oscillated with the circadian rhythm and whether NAMPT/visfatin, a critical enzyme in the NAD+ salvage pathway, was involved. Their results showed that CLOCK and SIRT1 regulate the expression of NAMPT/visfatin and are part of a circadian loop that links metabolism with the circadian clock.

So next time a 7-year old asks you the why-is-it-earlier-but-later question, give them the easy answer and tell them that obviously their CLOCK, SIRT1, or NAMPt/visfatin proteins must be out of whack.

Bethyl Laboratories has polyclonal antibodies against SIRT1 and NAMPT/visfatin for your studies of the circadian rhythm pathway. Additionally, antibodies against TIMELESS and TIPIN are also available.

Reference List

1. Nakahata, Y., S. Sahar, G. Astarita, M. Kaluzova, and P. Sassone-Corsi. 2009. Circadian Control of the NAD+ Salvage Pathway by CLOCK-SIRT1. Science.

HERC2, Don’t It Make My Brown Eyes Blue

2009-03-13T06:53:00.000-07:00

Does Crystal Gayle have blue or brown eyes? A Google search on this subject tells us that she is reported to have brown or hazel eyes (isn’t it great to have such critical information at our fingertips, thanks Google). But as the song says, when her man has found someone new, don’t it make her brown eyes blue? But what really does make someone’s eyes blue? And the even bigger question that some of you may have-especially if you are young and not a fan of 70’s country music- is who the heck is Crystal Gayle? You can Google that one on your own time, and as you probably guessed, losing your man or wo-man is not the cause of blue eyes. Human eye color is a multifactorial and complex hereditary trait. The variation in eye color from brown to green is a result of varying amounts of melanin in the iris, and blue eye color is the result of iris-specific melanin production being turned way down. Recently it has been found that blue eye color in European descendents probably resulted from a founder mutation that originated six to ten thousand years ago in the northwest part of the Black Sea region (1). The mutation has been discovered to be a single nucleotide polymorphism (SNP) fine-mapped to a short highly conserved sequence in intron 86 of the HERC2 (hect domain and RLD2) gene (1) (2). This SNP, rs1291382, is part of a consensus binding site for the helicase-like transcription factor (HLTF) also known as SMARCA3. Before this more definitive association of HERC2 to the blue-eye phenotype, the OCA2 (oculotaneous albinism 2) gene was primarily associated with the variation of eye color. As it turns out, OCA2 is critical in eye color determination, but it is the regulation of OCA2 by sequences mapped to rs1291382 in the neighboring HERC2 gene that are the main determiner of blue eye color. Eiberg et al have performed experiments in cell culture that have shown that the regulatory element containing the rs1291382 blue allele has inhibitory effects on OCA2 promoter activity. The study of regulatory elements and the factors that bind them will be a next step in understanding the complexity of eye color. Does SMARCA3 have a role? And what other factors might bind these sequences? The interesting subject of eye color also brings up the question of what are the exact cellular functions of the HERC2 and OCA2 gene products which remain uncharacterized, and why can’t I get that song out of my head?

For those interested in the cellular function of the HERC2 protein, antibodies to HERC2 are part of the list of new products for Bethyl Laboratories for the month of March. Also Bethyl Laboratories has two polyclonal products for the study of SMARCA3.

Reference List

1. Eiberg, H., J. Troelsen, M. Nielsen, A. Mikkelsen, J. Mengel-From, K. W. Kjaer, and L. Hansen. 2008. Blue eye color in humans may be caused by a perfectly associated founder mutation in a regulatory element located within the HERC2 gene inhibiting OCA2 expression. Hum.Genet. 123:177-187.
2. Sturm, R. A., D. L. Duffy, Z. Z. Zhao, F. P. Leite, M. S. Stark, N. K. Hayward, N. G. Martin, and G. W. Montgomery. 2008. A single SNP in an evolutionary conserved region within intron 86 of the HERC2 gene determines human blue-brown eye color. Am.J.Hum.Genet. 82:424-431.

New Polylconal Antibodies Released in February

2009-03-11T10:35:00.000-07:00

Antibodies qualified for IP/Western Blot: COBRA1 COX4 EEA1 EML4 eNOS EPS8 EpsinR FoxP3 GCF GEF-H1 HERC1 HERC2 HPK1 IDE MRP4 NELFA NELFE Notch1 ORC1 PARG1 RENT1 SOS1 SOS2 TAK1 VprBP WDR21A ZEB1

Antibodies qualified for IHC: ALMS1 CEP131/AZ1 CEP290 CEP78 CTF18 FOG1/ZFPM1 Jarid1B NUP153 WDFY3 WTAP ZNF326

Antibodies qualified for Flow: 53BP1 gamma-H2AX H2AX HdmX/MDM4 HIF1-alpha Histone H3 MDM4/HdmX MLL1 p53 PHD2 Phospho 53BP1 (S25) Phospho Histone H3 (S10) Raptor
Rictor

Tied to Telomeres

2009-02-17T19:08:00.000-08:00

So how do we tie a Spanish neuropathologist and cytologist named Santiago Ramón y Cajal who won a nobel prize in medicine in 1906 to an enzyme found to synthesize telomeric DNA and discovered only 20 years ago? Santiago Ramón y Cajal first observed “nucleolar accessory bodies” in the nuclei of neurons. Later, electron microscopist referred to these subnuclear organelles as “coiled bodies”; and much later these bodies were officially given the name “Cajal bodies” in honor of their accomplished discoverer. The exact function of Cajal bodies has proven to be enigmatic; almost as enigmatic as the exact pronunciation of “Cajal” (and so I will digress a moment for a quick lesson in the español).
If you weren’t aware that the Cajal body was named after a Spanish Nobel laureate you might want to anglicanize the word and pronounce the letter “J”, well, as the letter “J” is usually pronounced in English, (juh) . Unlike in English, the letter “J” in Spanish is pronounced like an “H”; so Cajal would be pronounced, kah-hahl′, as in the Spanish word everyone knows, cajones (kah-hō'-nēz) and not “kah-jahl” as in the English word Cajole (ke-jōl). But, back to the enigmatic function of Cajal bodies. For a while now, Cajal bodies have been implicated in ribonucleoprotein maturation, and it now looks like the coiled bodies represent a multifunctional organelle. Recently, Cajal bodies have been tied to the enzyme responsible for maintaining telomere length, telomerase. Several studies have shown that the subnuclear localization of telomerase is important to the regulation of its activity. This becomes more interesting as it has also been found that the localization of telomerase differs between normal and cancer cells; and this difference can be correlated with its activity as telomerase activity is normally absent from primary cells and is activated in cancer cells. The discovery that telomerase becomes re-activated in cancer cells, has led to an interest in understanding the details of this holoenzyme.
There are three known essential components of the active telomerase holoenzyme: dyskerin, TERT (telomerase RT), and TERC (telomerase RNA component). But in the January 30th issue of Science, Venteicher (Venteicher et al., 2009) and his collegues definitively report that they have discovered a fourth component critical to the activity of the telomerase holoenzyme. In search of proteins that interact with dyskerin, Venteicher et al. identified a previously uncharacterized WD-repeat protein known as WDR79 that appears to be instrumental to the localization of telomerase to Cajal bodies as well as its delivery to the telomeres during S-phase. Due to this assignment of function, the group refers to WDR79 as TCAB1 (telomerase Cajal body protein) and conclude that it is key to the regulation of telomerase localization. Now tied to telomerase and Cajal bodies, TCAB1 will be considered an important link to the understanding of telomerase regulation in cancer, and Señor Ramón y Cajal will continue to be associated with some of the most important findings of cellular biology today.

Check out Bethyl’s antibody against WDR79 (TCAB1) WDR79

Celebrity or Science?

2009-02-17T19:02:00.000-08:00

Those not familiar with miRNA (microRNA) and siRNA (short interfering RNA) might think that dicer, drosha, pasha, and loquacious are the members of a new up and coming pop music girl group. You know, Dicer is the tough one, Drosha- the mysterious one, Pasha- the snobby fashionable one, and loquacious- the outgoing talkative one. However as much as we need a new pop music girl group, these four individuals won’t serve to satisfy. Dicer, drosha, pasha, and loquacious are four proteins that may be destined for a much grander degree of celebrity status. Originally studied in flies and worms, these proteins have been discovered to be intimately involved in RNA interference, a process critical to the regulation of protein expression. Dr. Anil Sood of the University of Texas M.D. Anderson explains in simple terms how these proteins participate in the processing of mi- and siRNA (here). Homologs of dicer, drosha, and pasha have been identified in mammals and their study in human cells and tissues is advancing. Dr. Sood and his collegues have recently reported in the December 8, 2008 issue of the New England Journal of Medicine that dicer and drosha may have important implications for ovarian cancer (Merritt et al., 2008). In this study they suggest that low levels of both dicer and drosha are associated with shorter survival in ovarian cancer patients. In an M.D. Anderson Podcast, Dr. Sood explains what this means to the prognosis and treatment of cancer and what future directions lie ahead in the career of this class for proteins.

One of Bethyl’s newest polyclonal antibodies is against the human drosha protein.
Check it out on the Bethyl website: Drosha

New Polyclonal Antibodies Released in January

2009-02-08T18:38:00.000-08:00

Antibodies qualified for IP/Western Blot - CTF18, DjC21, DR1, Drosha, FOP, KANK1, LRWD1, SATB2, SMN1, UBE2O, UBE2T, UFD1, WDFY3, WDR70, WDR89, WNK3, ZC3H3, ZNF276, ZNF326, ZNF777.

Antibodies qualified for IHC - CSN5, ILK, IRS1, IRS2, MLH3, NRBF2, PTEN.

Antibodies to JUMONJI Proteins

2009-02-02T07:20:00.000-08:00

Accumulating evidence suggests that aberrant regulation of chromatin modification is important to tumorigenesis. Histone methylation is the posttranslational modification of histone amino acids by addition of methyl groups that regulates chromatin structure, transcription, and cellular epigenetic status. The jumonji (jmj) domain containing gene family of proteins has been discovered to play a role in chromatin regulation by influencing transcriptional activation, suppression, and demethylation. Two members of this protein family, JARID1A/RBP2 and JMJD2A, associate with the tumor suppressor protein pRb indicating their importance in transcriptional regulation and tumorigenesis. Furthermore, JMJD2C, also known as GASC1 (gene amplified in squamous cell carcinoma), has been found to be amplified in several espophageal squaumous carcinomas and inhibition of JMJD2C/GASC1 expression results in decreased cellular proliferation. These observations, associating jumonji-domain containing proteins with tumorigenesis, have identified histone demethylases an up and coming target of research for anti-cancer therapies.

Bethyl Laboratories' current portfolio of JUMONJI antibodies are JARID1A, JARID1B, JARID1C, JARID1D, JHDM1A, JMJD1A, JMJD1B, JMJD1C, JMJD2A, JMJD2B, JMJD2C.

Our complete profile of antibodies can be viewed at http://www.bethyl.com/.