Cell Signaling Technology

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

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
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 Blotting

Western Blotting

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 Blotting

Western Blotting

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 Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Bcl-2 (50E3) Rabbit mAb #2870.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Mcl-1 (D35A5) Rabbit mAb #5453.

Description

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.

Background

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).

  1. Cory, S. et al. (2003) Oncogene 22, 8590-607.
  2. Antonsson, B. and Martinou, J.C. (2000) Exp Cell Res 256, 50-7.
  3. Sharpe, J.C. et al. (2004) Biochim Biophys Acta 1644, 107-13.
  4. Korsmeyer, S.J. et al. (1993) Semin Cancer Biol 4, 327-32.
  5. Bouillet, P. and Strasser, A. (2002) J Cell Sci 115, 1567-74.
  6. Maundrell, K. et al. (1997) J. Biol. Chem. 272, 25238-25242.
  7. Yamamoto, K. et al. (1999) Mol. Cell Biol. 19, 8469-8478.
  8. Ling, Y. H. et al. (1998) J. Biol. Chem. 273, 18984-18991.
  9. Huang, S.J. and Cidlowski, J.A. (2002) FASEB 16, 825-832.
  10. Deng, X. et al. (2001) J. Biol. Chem. 276, 23681-23688.

Application References

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Protocols

* Product-specific protocol.

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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