Product Pathways - Apoptosis
Pro-Survival Bcl-2 Family Antibody Sampler Kit #9941
|Kit Includes||Quantity||Applications||Reactivity||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 IgG|
|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|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey C=Chicken B=Bovine Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
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.
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.
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.