Bad Antibody

9292:

Immunoprecipitation , Western Blotting

Bad Antibody detects endogenous levels of total Bad protein. The antibody does not cross-react with related proteins.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser112 of mouse Bad. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence^® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence^® Bad siRNA I (+), using Bad Antibody and p42 MAPK Antibody #9108. The Bad antibody confirms silencing of Bad expression, while the p42 MAPK antibody is used to control for loading and specificity of Bad siRNA.

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence^® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence^® Bad siRNA II (+), using Bad Antibody and α-Tubulin (11H10) Rabbit mAb #2125. The Bad antibody confirms silencing of Bad expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Bad siRNA.

Western Blotting

Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Bad Antibody (right) or Phospho-Bad (Ser112) Antibody #9291 (left).

Western Blotting

Western blot analysis of extracts from 293 cells transfected with Wild-type Bad, Bad (Ser112A), Bad (S136A) or Bad (S112A/S136A), untreated, TPA-treated or forskolin-treated, using Phospho-Bad (Ser112) Antibody #9291 (top), Phospho-Bad (Ser136) Antibody #9295 (middle) or Bad Antibody (bottom).

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.

• 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 co-activator. Mol. Cell 6, 395-407. Applications: Western Blotting
• Yu, C. et al. (2003) The lethal effects of pharmacological cyclin-dependent kinase inhibitors in human leukemia cells proceed through a phosphatidylinositol 3-kinase/Akt-dependent process. Cancer Res. 63, 1822-1833. 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
• Gómez-Abad, C. et al. (2011) Blood 118, 5517-27. 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 !

• 5284 Phospho-Bad (Ser112) (40A9) Rabbit mAb
• 5286 Phospho-Bad (Ser136) (185D10) Rabbit mAb
• 9293 Bad Control Proteins
• 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

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