Pro-Survival Bcl-2 Family Antibody Sampler Kit II #17229
Product Information
Kit Usage Information
Protocols
- 2724: Western Blotting
- 2764: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence*, Flow
- 2827: Western Blotting, Flow
- 4223: Western Blotting, Immunoprecipitation (Agarose)
- 7074: Western Blotting
- 14093: Western Blotting, Immunoprecipitation (Magnetic)
- 14765: Western Blotting, Immunoprecipitation (Magnetic)
- 94296: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence, Flow
Product Description
Specificity / Sensitivity
Each antibody in the Pro-Survival Bcl-2 Family Antibody Sampler Kit II recognizes endogenous levels of its specific target. The antibodies do not cross-react with other Bcl-2 family members. A1/Bfl-1 (D1A1C) Rabbit mAb may cross-react with an unknown protein at 50 and 130 kDa in some cell lines. Phospho-Bcl-2 (Ser70) (5H2) Rabbit mAb detects endogenous of human Bcl-2 only when phosphorylated at Ser70. Phospho-Mcl-1 (Thr163) (D5M9D) Rabbit mAb recongizes endogenous levels of Mcl-1 only when phosphorylated at Thr163. This antibody may also cross-react with an unidentified protein at 70 kDa in some cell lines.
Source / Purification
Rabbit monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Gly47 of human Bcl-2, Asp61 of human Bcl-xL, Pro60 of mouse Mcl-1, Gly29 of human A1/Bfl-1, and Ala39 of human Bcl-w. Phospho-specific rabbit monoclonal antibodies are produced by immunizing animlars with synthetic phospho-peptides correspoding to residues surrounding Ser70 of human Bcl-2 and Thr163 of human Mcl-1.
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).
Mcl-1 is phosphorylated in response to treatment with phorbol ester, microtubule-damaging agents, oxidative stress, and cytokine withdrawal (11-14). Phosphorylation at Thr163, the conserved MAP kinase/ERK site located within the PEST region, slows Mcl-1 protein turnover (13) but may prime the GSK-3 mediated phosphorylation at Ser159 that leads to Mcl-1 destabilization (14).
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- Ling, Y.H. et al. (1998) J Biol Chem 273, 18984-91.
- Huang, S.T. and Cidlowski, J.A. (2002) FASEB J 16, 825-32.
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