Catalog # RK6080

Raf Phospho-Regulation Antibody Kit

Antibody Sampler Kit

DATASHEET  




$395

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 in B-Raf is required for growth factor activation and kinase activity.

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

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).

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-B-Raf (N-terminus) (lanes 1 & 2) or anti-B-Raf (Ser-446) (lanes 3 & 4).



Mouse monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store all 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.



*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:

CATALOG #DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
A-Raf (N-terminal region) Mouse mAb50 μlWB, EHu, Rt, Ms68
B-Raf (N-terminus) Rabbit pAb50 μlWB, EHu, Rt, Ms95
B-Raf (S446)[C-Raf (S338)/A-Raf (S299)], phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms95
C-Raf (C-terminus) Rabbit pAb50 μlWB, EHu, Rt, Ms74
C-Raf (N-terminal region) Mouse mAb50 μlWB, EHu, Rt, Ms74
C-Raf (S471)[B-Raf (S579)/A-Raf (S432)], phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms74
KIT SUMMARY

The Raf phospho-regulation antibody sampler kit can be used to examine phosphorylation of A-Raf, B-Raf, and C-Raf. The anti-B-Raf (Ser-446) detects site-specific phosphorylation of B-Raf, as well as the conserved site in C-Raf (Ser-338) and A-Raf (Ser-299). Similarly, the anti-C-Raf (Ser-471) detects site-specific phosphorylation in C-Raf, as well as in B-Raf (Ser-579) and A-Raf (Ser-432). The kit also includes antibodies to monitor total expression levels of A-Raf, B-Raf, and C-Raf.