Product Pathways - Apoptosis
Phospho-Bim (Ser77) (D4H12) Rabbit mAb #12433
|12433S||100 µl (10 western blots)||---||In Stock||---|
|12433||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse||Endogenous||26||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation
Species predicted to react based on 100% sequence homology: Monkey.
Specificity / Sensitivity
Phospho-Bim (Ser77) (D4H12) Rabbit mAb recognizes endogenous levels of Bim protein only when phosphorylated at Ser77.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser77 of human Bim protein.
Western blot analysis of extracts from Raji and MCF7 cells, untreated (-) or treated with TPA #4174 (200 nM, 30 min; +), using Phospho-Bim (Ser77) (D4H12) Rabbit mAb (upper) or Bim (C34C5) Rabbit mAb #2933 (lower).
Immunoprecipitation of Phospho-Bim (Ser77) from extracts of MCF7 cells treated with TPA #4174 (200 nM, 30 min), using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-Bim (Ser77) (D4H12) Rabbit mAb (lane 3). Lane 1 is 10% input. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used to avoid recognition of rabbit IgG heavy and light chains by western blot.
Bim/Bod is a pro-apoptotic protein belonging to the BH3-only group of Bcl-2 family members including Bad, Bid, Bik, Hrk, and Noxa that contain a BH3 domain but lack other conserved BH1 or BH2 domains (1,2). Bim induces apoptosis by binding to and antagonizing anti-apoptotic members of the Bcl-2 family. Interactions have been observed with Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bfl-1, and BHRF-1 (1,2). Bim functions in regulating apoptosis associated with thymocyte negative selection and following growth factor withdrawal, during which Bim expression is elevated (3-6). Three major isoforms of Bim are generated by alternative splicing: BimEL, BimL, and BimS (1). The shortest form, BimS, is the most cytotoxic and is generally only transiently expressed during apoptosis. The BimEL and BimL isoforms may be sequestered to the dynein motor complex through an interaction with the dynein light chain and released from this complex during apoptosis (7). Apoptotic activity of these longer isoforms may be regulated by phosphorylation (8,9). Environmental stress triggers Bim phosphorylation by JNK and results in its dissociation from the dynein complex and increased apoptotic activity.
MAP kinase dependent phosphorylation of Bim at multiple sites, including Ser55, Ser65, and Ser73 in mouse (Ser59, Ser69, and Ser77 in human), can promote proteasomal degradation of Bim and inhibition of apoptosis (10).
- O'Connor, L. et al. (1998) EMBO J 17, 384-95.
- Hsu, S.Y. et al. (1998) Mol Endocrinol 12, 1432-40.
- Bouillet, P. et al. (2002) Nature 415, 922-6.
- Whitfield, J. et al. (2001) Neuron 29, 629-43.
- Dijkers, P.F. et al. (2000) Curr Biol 10, 1201-4.
- Ley, R. et al. (2003) J Biol Chem 278, 18811-6.
- Puthalakath, H. et al. (1999) Mol Cell 3, 287-96.
- Lei, K. and Davis, R.J. (2003) Proc Natl Acad Sci U S A 100, 2432-7.
- Putcha, G.V. et al. (2003) Neuron 38, 899-914.
- Hübner, A. et al. (2008) Mol Cell 30, 415-25.
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