Phospho-Bim (Ser69) (D7E11) Ra

4585:

Immunoprecipitation , Western Blotting

Phospho-Bim (Ser69) (D7E11) Rabbit mAb detects endogenous levels of Bim protein only when phosphorylated at Ser69.

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser69 of human Bim protein.

Western Blotting

Western blot analysis of extracts from Raji cells, untreated or TPA-treated for 30 min, using Phospho-Bim (Ser69) (D7E11) Rabbit mAb (upper) or total Bim (C34C5) Rabbit mAb #2933 (lower).

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: Bim_EL , Bim_L and Bim_S (1). The shortest form, Bim_S , is the most cytotoxic and is generally only transiently expressed during apoptosis. The Bim_EL and Bim_L 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.

ERK 1/2-dependent phosphorylation of Bim_EL at Ser69 (Ser65 in mouse and rat) in response to growth factor stimulation can promote its proteasome-mediated degradation and enhance cell survival (6,10,11).

1. O'Connor, L. et al. (1998) EMBO J 17, 384-95.
2. Hsu, S.Y. et al. (1998) Mol Endocrinol 12, 1432-40.
3. Bouillet, P. et al. (2002) Nature 415, 922-6.
4. Whitfield, J. et al. (2001) Neuron 29, 629-43.
5. Dijkers, P.F. et al. (2000) Curr Biol 10, 1201-4.
6. Ley, R. et al. (2003) J Biol Chem 278, 18811-6.
7. Puthalakath, H. et al. (1999) Mol Cell 3, 287-96.
8. Lei, K. and Davis, R.J. (2003) Proc Natl Acad Sci U S A 100, 2432-7.
9. Putcha, G.V. et al. (2003) Neuron 38, 899-914.
10. Luciano, F. et al. (2003) Oncogene 22, 6785-93.
11. Marani, M. et al. (2004) Oncogene 23, 2431-41.

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• 4581 Phospho-Bim (Ser69) Antibody
• 4550 Phospho-Bim (Ser55) Antibody
• 2933 Bim (C34C5) Rabbit mAb
• 2819 Bim Antibody
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked 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.