Product Pathways - Apoptosis
Pro-Survival Bcl-2 Family Antibody Sampler Kit #9941
|9941S||1 Kit (5 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb #2827||40 µl||W, IF-IC, F||H||28||Rabbit IgG|
|Phospho-Bcl-2 (Thr56) Antibody (Human Specific) #2875||40 µl||W||H||28||Rabbit|
|Bcl-2 (50E3) Rabbit mAb #2870||40 µl||W, IP||H, M, R||Mk, C, B, Dg||26||Rabbit|
|Bcl-xL (54H6) Rabbit mAb #2764||40 µl||W, IP, IHC-P, IHC-F, IF-IC, F||H, M, R, Mk||30||Rabbit IgG|
|Mcl-1 (D35A5) Rabbit mAb #5453||40 µl||W||H, M, Mk||B||40 (human), 35 (mouse)||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IHC-F=Immunohistochemistry (Frozen)
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey
Western blot analysis of extracts from Jurkat and HeLa (human), COS (monkey), NIH/3T3 and L929 (mouse), and PC12 and C6 (rat) cells, using Bcl-xL (54H6) Rabbit mAb #2764.
Western blot analysis of extracts from THP-1 cells, untreated or nocadazole-treated (1.0 µg/mL) overnight, using Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb #2827 (upper), Phospho-Bcl-2 (Thr56) Antibody (Human Specific) #2875 (middle) and Bcl-2 (50E3) Rabbit mAb #2870 (lower).
Western blot analysis of extracts from various cell types using Bcl-2 (50E3) Rabbit mAb #2870.
The Pro-Survival Bcl-2 Family Antibody Sampler Kit provides an economical means to examine several members of the Bcl-2 family. The kit contains enough primary and secondary antibodies to perform four western blot experiments.
Specificity / Sensitivity
Each antibody in the Pro-Survival Bcl-2 Family Antibody Sampler Kit recognizes only its specific target. The antibodies do not cross-react with other Bcl-2 family members. Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb detects endogenous levels of human Bcl-2 only when phosphorylated at Ser70. Phospho-Bcl-2 (Thr56) Antibody (Human Specific) detects endogenous levels of human Bcl-2 only when phosphorylated at Thr56.
Source / Purification
Total Mcl-1, Bcl-xL, and Bcl-2 monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu210 of human Mcl-1, Asp61 of human Bcl-xL, and residues at the carboxy terminus of human Bcl-2 alpha. Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser70 of human Bcl-2. Phospho-Bcl-2 (Thr56) Antibody (Human Specific) is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr56 of human Bcl-2. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
The Bcl-2 family consists of a number of evolutionarily conserved proteins containing Bcl-2 homology domains (BH) that regulate apoptosis through control of mitochondrial membrane permeability and release of cytochrome c (1-3). Four BH domains have been identified (BH1-4) that mediate protein interactions. The family can be separated into three groups based upon function and sequence homology: pro-survival members include Bcl-2, Bcl-xL, Mcl-1, A1 and Bcl-w; pro-apoptotic proteins include Bax, Bak and Bok, and "BH3 only" proteins Bad, Bik, Bid, Puma, Bim, Bmf, Noxa and Hrk. Interactions between death-promoting and death-suppressing Bcl-2 family members has led to a rheostat model in which the ratio of pro-apoptotic and anti-apoptotic proteins controls cell fate (4). Thus, pro-survival members exert their behavior by binding to and antagonizing death-promoting members. In general, the "BH3-only members" can bind to and antagonize the pro-survival proteins leading to increased apoptosis (5). While some redundancy of this system likely exists, tissue specificity, transcriptional and post-translational regulation of many of these family members can account for distinct physiological roles.
Several phosphorylation sites have been identified within Bcl-2 including Thr56, Ser70, Thr74 and Ser87 (6). These phosphorylation sites may be targets of the ASK1/MKK7/JNK1 pathway, and phosphorylation of Bcl-2 may be a marker for mitotic events (7,8). Mutation of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity during glucocorticoid-induced apoptosis of T lymphocytes (9). Interleukin 3 and JNK-induced Bcl-2 phosphorylation at Ser70 may be required for its enhanced antiapoptotic functions (10).
- Cory, S. et al. (2003) Oncogene 22, 8590-607.
- Antonsson, B. and Martinou, J.C. (2000) Exp Cell Res 256, 50-7.
- Sharpe, J.C. et al. (2004) Biochim Biophys Acta 1644, 107-13.
- Korsmeyer, S.J. et al. (1993) Semin Cancer Biol 4, 327-32.
- Bouillet, P. and Strasser, A. (2002) J Cell Sci 115, 1567-74.
- Maundrell, K. et al. (1997) J. Biol. Chem. 272, 25238-25242.
- Yamamoto, K. et al. (1999) Mol. Cell Biol. 19, 8469-8478.
- Ling, Y. H. et al. (1998) J. Biol. Chem. 273, 18984-18991.
- Huang, S.J. and Cidlowski, J.A. (2002) FASEB 16, 825-832.
- Deng, X. et al. (2001) J. Biol. Chem. 276, 23681-23688.
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* Product-specific protocol.
- 9942 Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit
- 4223 Bcl-2 (D55G8) Rabbit mAb (Human Specific)
- 3498 Bcl-2 (D17C4) Rabbit mAb (Mouse Preferred)
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.