Cell Signaling Technology

Product Pathways - Apoptosis

Phospho-Bad (Ser136) (D25H8) Rabbit mAb #4366

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M Mk (R) Endogenous 23 Rabbit IgG

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

Protocols

Specificity / Sensitivity

Phospho-Bad (Ser136) (D25H8) Rabbit mAb detects endogenous levels of Bad only when phosphorylated at Ser136 (equivalent to Ser99 in human and Ser137 in rat).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser136 of mouse Bad protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from ACHN cells, untreated or treated with Human Epidermal Growth Factor #8916 (100 ng/ml, 30 min), and C2C12 cells, untreated or insulin-treated (100 nM, 30 min), using Phospho-Bad (Ser136) (D25H8) Rabbit mAb (upper) or Bad (D24A9) Rabbit mAb #9239 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from COS-7 cells, treated with Human Epidermal Growth Factor #8916 (100 ng/ml, 30 min) in the presence or absence of calf intestinal phosphatase (CIP) plus lambda-phosphatase, using Phospho-Bad (Ser136) (D25H8) Rabbit mAb (upper) or Bad (D24A9) Rabbit mAb #9239 (lower).

Western Blotting

Western Blotting

Western blot analysis of lysates containing Bad Control Proteins #9293, comprised of nonphosphorylated and phosphorylated Bad peptides, using Phospho-Bad (Ser136) (D25H8) Rabbit mAb.


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 at 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

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Companion Products

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

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