Phospho-c-Raf (Ser259) AntibodyProduct information
Product Pathways - MAPK Signaling
Phospho-c-Raf (Ser259) Antibody #9421
|9421S||100 µl (10 western blots)||---||In Stock||---|
|9421P||40 µl (4 western blots)||---||In Stock||---|
|9421||carrier free and custom formulation / quantity||email request|
|W||1:1000||Human, Mouse, Rat, Monkey, Xenopus||Endogenous||74||Rabbit|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation
Species predicted to react based on 100% sequence homology: Chicken.
Specificity / Sensitivity
Phospho-c-Raf (Ser259) Antibody detects endogenous levels of c-Raf only when phosphorylated at Ser259.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser259 of human c-Raf. Antibodies are purified by protein A and peptide affinity chromatography.
Site specificity of Phospho-c-Raf (Ser259) Antibody: Western blot analysis of recombinant Myc-tagged c-Raf protein, wild-type (lanes 1 and 3) and S259A mutant (lanes 2 and 4), using Phospho-Raf (Ser259) Antibody or a Myc antibody. (Provided by Dr. Guri Tzivion, Massachusetts General Hospital.)
A-Raf, B-Raf, and c-Raf (Raf-1) are the main effectors recruited by GTP-bound Ras to activate the MEK-MAP kinase pathway (1). Activation of c-Raf is the best understood and involves phosphorylation at multiple activating sites including Ser338, Tyr341, Thr491, Ser494, Ser497, and Ser499 (2). p21-activated protein kinase (PAK) has been shown to phosphorylate c-Raf at Ser338, and the Src family phosphorylates Tyr341 to induce c-Raf activity (3,4). Ser338 of c-Raf corresponds to similar sites in A-Raf (Ser299) and B-Raf (Ser445), although this site is constitutively phosphorylated in B-Raf (5). Inhibitory 14-3-3 binding sites on c-Raf (Ser259 and Ser621) can be phosphorylated by Akt and AMPK, respectively (6,7). While A-Raf, B-Raf, and c-Raf are similar in sequence and function, differential regulation has been observed (8). Of particular interest, B-Raf contains three consensus Akt phosphorylation sites (Ser364, Ser428, and Thr439) and lacks a site equivalent to Tyr341 of c-Raf (8,9). Research studies have shown that the B-Raf mutation V600E results in elevated kinase activity and is commonly found in malignant melanoma (10). Six residues of c-Raf (Ser29, Ser43, Ser289, Ser296, Ser301, and Ser642) become hyperphosphorylated in a manner consistent with c-Raf inactivation. The hyperphosphorylation of these six sites is dependent on downstream MEK signaling and renders c-Raf unresponsive to subsequent activation events (11).
- Avruch, J. et al. (1994) Trends Biochem Sci 19, 279-83.
- Chong, H. et al. (2001) EMBO J 20, 3716-27.
- King, A.J. et al. (1998) Nature 396, 180-3.
- Fabian, J.R. et al. (1993) Mol Cell Biol 13, 7170-9.
- Mason, C.S. et al. (1999) EMBO J 18, 2137-48.
- Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.
- Sprenkle, A.B. et al. (1997) FEBS Lett 403, 254-8.
- Marais, R. et al. (1997) J Biol Chem 272, 4378-83.
- Guan, K.L. et al. (2000) J Biol Chem 275, 27354-9.
- Davies, H. et al. (2002) Nature 417, 949-54.
- Dougherty, M.K. et al. (2005) Mol Cell 17, 215-24.
- Cieslik, K. et al. (2001) J. Biol. Chem. 276, 1211-1219. Applications: Western Blotting.
- Weber, S. M. et al. (2002) J. Immunol. 168, 5303-5309. Applications: Western Blotting.
- Tap, W.D. et al. (2010) Neoplasia 12, 637-49. 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!
- 4432 A-Raf Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7727 Biotinylated Protein Ladder Detection Pack
- 4431 Phospho-A-Raf (Ser299) Antibody
- 2696 Phospho-B-Raf (Ser445) Antibody
- 9427 Phospho-c-Raf (Ser338) (56A6) Rabbit mAb
- 6883 SignalFire™ ECL Reagent
- 12757 SignalFire™ Elite ECL Reagent
- 12630 SignalFire™ Plus ECL Reagent
- 53745 c-Raf (D4B3J) Rabbit mAb
- 9422 c-Raf Antibody
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.