Phospho-Bad (Ser155) Antibody

9297:

Western Blotting

Phospho-Bad (Ser155) Antibody detects transfected levels of Bad only when phosphorylated at Ser155. This antibody does not detect Bad phosphorylated at other sites or related family members.

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser155 of mouse Bad. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of cell extracts from 293 cells transfected with Wild-type Bad, Bad (S112A/S136A), Bad (S155A) and treated with forskolin (30 µM), using Phospho-Bad (Ser155) Antibody (upper) and Bad Antibody #9292 (lower).

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.

• Tan, Y. et al. (2000) BAD Ser-155 phosphorylation regulates BAD/Bcl-XL interaction and cell survival. J. Biol. Chem. 275, 25865-25869. Applications: Western Blotting
• Masters, S. et al. (2001) 14-3-3 inhibits Bad-induced cell death through interaction with serine-136. Mol. Pharmacol. 60 (6), 1325-1331. Applications: Western Blotting
• Yusta, B. et al. (2002) Glucagon-like peptide-2 receptor activation engages Bad and glycogen synthase kinase-3 in a protein kinase A-dependent manner and prevents apoptosis following inhibition of phosphatidylinositol 3-kinase. J. Biol. Chem. 277 (28), 24896-24906. Applications: Western Blotting
• Saito, A. et al. (2003) Overexpression of copper/zinc superoxide dismutase in transgenic mice protects against neuronal cell death after transient focal ischemia by blocking activation of the Bad cell death signaling pathway. J. Neurosci. 23, 1710-1718. Applications: Western Blotting

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• 5284 Phospho-Bad (Ser112) (40A9) Rabbit mAb
• 5286 Phospho-Bad (Ser136) (185D10) Rabbit mAb
• 9292 Bad Antibody
• 9296 Phospho-Bad (Ser112) (7E11) Mouse mAb
• 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.