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).
The B-Raf mutation V600E results in elevated kinase activity and is commonly found in malignant melanoma.
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 Avruch, J.
et al.
(1994) Trends Biochem.
Sci.
19, 279-283.
Chong, H.
et al.
(2001) EMBO J.
20, 3716-3727.
King, A.J.
et al.
(1998) Nature 396, 180-183.
Fabian, J.R.
et al.
(1993) Mol.
Cell Biol.
13, 7170-7179.
Applications:Suitable for use in Western Blot.
Other applications not tested.
Recommended Dilution:Western Blot: 1:500-1:1000Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：100ul
Host：rabbit
Source Antibody：rat
Grade：Affinity Purified
Purity：Purified by immunoaffinity chromatography.
Form：Supplied as a liquid in PBS, pH 7.4, 150mM sodium chloride, 0.02% sodium azide, 50% glycerol.
Specificity：Rat
Isotype：IgG
Calc Applications Abbrev：WB
Calc Crossreactivity：Rt
Immunogen：Peptide corresponding to the sequence around aa641-645(T-S-P-R-L ) derived from rat Raf-1.
