Catalog # RP2901

C-Raf (S471)/B-Raf (S579)/A-Raf (S432), phospho-Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 74 kDa

The Ras-Raf-MAP kinase signaling pathway is involved in control of cell proliferation and differentiation. The Raf kinase family includes A-Raf, B-Raf, and C-Raf. Each family member has three highly conserved regions (CR1-3). The N-terminal CR1 contains the Ras-GTP-binding domain. The CR2 contains a negative regulatory serine residue (C-Raf (S259)/B-Raf(S365)) that may bind 14-3-3 proteins. The CR3 is the catalytic domain that contains phosphorylation sites for Raf-regulating enzymes within two segments, the N-region and the activation segment. Activation of C-Raf involves phosphorylation at many sites including Ser-338, Tyr-341, and Ser-471. The latter site is phosphorylated after EGF stimulation and may be important for MEK interaction in both C-Raf and A-Raf. In B-Raf, multiple phosphorylation sites have been identified, but their specific roles are uncertain. Phosphorylation of Ser-446 may prime B-Raf for activation, and Ser-446 and/or Ser-447 phosphorylation may be critical for B-Raf biological activity during PC12 differentiation. Ser-579 is required for growth factor activation and kinase activity.

References

Baljuls, A. et al. (2008) J Biol Chem 283(40):27239.
Brummer, T. et al. (2006) Oncogene 25(47):6262.
Karbowniczek, M. et al. (2006) J Biol Chem 281(35):25447.
Zhu, J. et al. (2005) Mol. Biol. Cell 16:4733.
Wilhelm, S.M. et al. (2004) Cancer Res 64:7099.
Mason, C.S. et al. (1999) EMBOJ 18(8):2137.

Western blot of human Jurkat cells treated with calyculin A (100 nM) for 30 min. The blots were untreated (lanes 1 & 3) or treated (lanes 2 & 4) with lambda phosphatase and probed with anti-C-Raf (N-terminal region) (lanes 1 & 2) or anti-C-Raf (Ser-471) (lanes 3 & 4).

A synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding Ser-471 in human C-Raf. This sequence has high homology with similar regions in rat and mouse C-Raf, and has high homology to the conserved sites in B-Raf (Ser-579) and A-Raf (Ser-432).

*For more information, see UniProt Accession P04049

Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody was cross-adsorbed to phospho-C-Raf (Ser-338) peptide before affinity purification using phospho-C-Raf (Ser-471) peptide. The purified antibody detects a band at 74 kDa* corresponding to C-Raf in western blots of human Jurkat cells treated with Calyculin A, but is not observed after lambda phosphatase treatment.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: