Product Pathways - Apoptosis
Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit #9942
|9942S||1 Kit (8 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Bad (D24A9) Rabbit mAb #9239||40 µl||W||H, M, R, Mk||23||Rabbit IgG|
|Phospho-Bad (Ser112) (40A9) Rabbit mAb #5284||40 µl||W, IHC-P, F||H, M, R, Mk||23||Rabbit IgG|
|Bax (D2E11) Rabbit mAb #5023||40 µl||W, IP, IHC-P||H||20||Rabbit IgG|
|Bik Antibody #4592||40 µl||W, IHC-P||H||20||Rabbit|
|Bim (C34C5) Rabbit mAb #2933||40 µl||W, IP, IHC-P, IF-IC, F||H, M, R||Mk, B, Dg||12, 15, 23||Rabbit|
|BID Antibody (Human Specific) #2002||40 µl||W, IP||H||15, 22||Rabbit|
|Bak (D2D3) Rabbit mAb #6947||40 µl||W, IP, IHC-P||H, Mk||25||Rabbit IgG|
|Puma Antibody #4976||40 µl||W||H||Mk||23||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey
Western blot analysis of extracts from Jurkat cells, untreated or etoposide-treated (25 µM), and RS4;11 cells, untreated or okadaic acid-treated (1 µM), using BID Antibody (Human Specific) Antibody #2002.
Western blot analysis of extracts from Raji, A20 and RL-7 cells using Bim (C34C5) Rabbit mAb #2933.
Western blot analysis of extracts from Raji and Ramos cell lines using Bik Antibody #4592.
Western blot analysis of extracts from various cell lines using Bax (D2E11) Rabbit mAb #5023. Brimmel et al. demonstated that Jurkat cells lack Bax protein expression (10).
Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Phospho-Bad (Ser112) (40A9) Rabbit mAb #5284 (upper) or Bad Antibody #9292 (lower).
Western blot analysis of extracts from various cell lines using Bak (D2D3) Rabbit mAb #6947.
The Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit provides an economical means to examine several members of the Bcl-2 family and their activation status. The kit contains enough primary and secondary antibodies to perform four Western blot experiments per primary antibody.
Specificity / Sensitivity
Each antibody in the Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit recognizes only its specific target. The antibodies do not cross-react with other Bcl-2 family members. Phospho-Bad (Ser112) (40A9) Rabbit mAb detects endogenous levels of Bad only when phosphorylated at Ser112 (mouse), Ser75 (human), or Ser113 (rat).
Source / Purification
Phospho-Bad (Ser112) (40A9) Rabbit mAb is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser112 of mouse Bad. Total Bad, Bax, Bim, and Bak monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro102 of human Bad, Leu45 of human Bax, Pro25 of human Bim, or Gly82 of human Bak. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the amino-terminus of human Bik; residues surrounding the cleavage site of human BID; or the carboxy terminus of human Puma. 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.
Bad is a pro-apoptotic member of the Bcl-2 family that can displace Bax from binding to Bcl-2 and Bcl-xL, resulting in cell death (6,7). Survival factors such as IL-3 can inhibit the apoptotic activity of Bad by activating intracellular signaling pathways that result in the phosphorylation of Bad at Ser112 and Ser136 (7). Phosphorylation at these sites results in the binding of Bad to 14-3-3 proteins and the inhibition of Bad binding to Bcl-2 and Bcl-xL (7). Akt has been shown to promote cell survival via its ability to phosphorylate Bad at Ser136 (8,9). Ser112 has been shown to be the substrate in vivo and in vitro of p90RSK (10,11) and mitochondria-anchored PKA (12).
- 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.
- Yang, E. et al. (1995) Cell 80, 285-291.
- Zha, J. et al. (1996) Cell 87, 619-628.
- Datta, S. R. et al. (1997) Cell 91, 231-241.
- Peso, L. et al. (1997) Science 278, 687-689.
- Bonni, A. et al. (1999) Science 286, 1358-1362.
- Tan, Y. et al. (1999) J. Bio. Chem. 274, 34859-34867.
- Harada, H. et al. (1999) Mol. Cell 3, 413-422.
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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.