Raf Family, Antibody Sampler Kit, BioAssay(TM)

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 (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). Kit Includes: Quantity Applications Reactivity MW (kD) (1) Rabbit x Phospho-A-Raf (Ser299) Antibody 40ul WB Human Mouse (Rat) 68kD Rabbit (2) Rabbit x A-Raf Antibody 40ul WB IP Human Mouse Rat 68kD (3) Rabbit x Phospho-c-Raf (Ser338) (mAb) 40ul WB Human Mouse Rat Monkey 74kD (4) Rabbit x Phospho-c-Raf (Ser289/296/301) 40ul WB Human Mouse 74kD (5) Rabbit x Phospho-c-Raf (Ser259) 40ul WB IP Human Mouse Rat Xenopus (Chicken) 74kD (6) Rabbit x c-Raf Antibody 40ul WB IP Human Mouse Rat 65 to 75kD (7) Rabbit x Phospho-B-Raf (Ser445) 40ul WB Human Mouse Rat Monkey (Chicken) 86kD (8) Mouse x B-Raf (mAb) 40ul WB IP FC Human Mouse Rat Monkey 86kD