Product Pathways - Apoptosis / Autophagy

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Phospho-Bad (Ser136) Antibody #9295

Applications	Reactivity	Sensitivity	Source
W	M (H) (R)	Transfected Only	Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

9295:

Western Blotting

Specificity / Sensitivity

Phospho-Bad (Ser136) Antibody detects transfected levels of Bad only when phosphorylated at Ser136. This antibody does not detect Bad phosphorylated at other sites, nor does it detect related family members.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser136 of mouse Bad. Antibodies are purified by protein A and peptide affinity chromatography.

Background

Bad is a proapoptotic member of the Bcl-2 family that promotes cell death by displacing Bax from binding to Bcl-2 and Bcl-xL (1,2). Survival factors, such as IL-3, inhibit the apoptotic activity of Bad by activating intracellular signaling pathways that result in the phosphorylation of Bad at Ser112 and Ser136 (2). Phosphorylation at these sites promotes binding of Bad to 14-3-3 proteins to prevent an association between Bad with Bcl-2 and Bcl-xl (2). Akt phosphorylates Bad at Ser136 to promote cell survival (3,4). Bad is phosphorylated at Ser112 both in vivo and in vitro by p90RSK (5,6) and mitochondria-anchored PKA (7). Phosphorylation of Ser155 in the BH3 domain by PKA plays a critical role in blocking the dimerization of Bad and Bcl-xL (8-10).

1. Yang, E. et al. (1995) Cell 80, 285-291.
2. Zha, J. et al. (1996) Cell 87, 619-628.
3. Datta, S.R. et al. (1997) Cell 91, 231-241.
4. Peso, L. et al. (1997) Science 278, 687-689.
5. Bonni, A. et al. (1999) Science 286, 1358-1362.
6. Tan, Y. et al. (1999) J. Biol. Chem. 274, 34859-34867.
7. Harada, H. et al. (1999) Mol. Cell 3, 413-422.
8. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
9. Lizcano, J. et al. (2000) Biochem. J. 349, 547-557.
10. Datta, S. et al. (2000) Mol. Cell 6, 41-51.

Application References

• Jamieson, C.A. and Yamamoto, K.R. (2000) Crosstalk pathway for inhibition of glucocorticoid-induced apoptosis by T cell receptor signaling. Proc. Natl. Acad. Sci. USA 97, 7319-7324. Applications: Western Blotting
• Laine, J. et al. (2000) The protooncogene TCL1 is an Akt kinase coactivator. Mol. Cell 6, 395-407. Applications: Western Blotting
• Yang, C. C. et al. (2003) Bcl-xL mediates a survival mechanism independent of the phosphoinositide 3-kinase/Akt pathway in prostate cancer cells. J. Biol. Chem. 278, 25872-25878. Applications: Western Blotting
• Rusinol, A. E. et al. (2004) AKT/protein kinase B regulation of BCL family members during oxysterol-induced apoptosis. J. Biol. Chem. 279, 1392-1399. Applications: Western Blotting
• Tan, Y. et al. (1999) p90(RSK) blocks bad-mediated cell death via a protein kinase C-dependent pathway. J. Biol. Chem. 274, 34859-34867. Applications: Western Blotting
• Gray, M.J. et al. (2008) J Natl Cancer Inst 100, 109-20. Applications: Western Blotting
• Fujita, H. et al. (2005) Biochem Pharmacol 69, 1773-84. Applications: Western Blotting

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

• 5284 Phospho-Bad (Ser112) (40A9) Rabbit mAb
• 5286 Phospho-Bad (Ser136) (185D10) Rabbit mAb
• 9292 Bad Antibody
• 9293 Bad Control Proteins
• 9296 Phospho-Bad (Ser112) (7E11) Mouse mAb
• 9297 Phospho-Bad (Ser155) Antibody
• 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

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.