CST, 2696S, Phospho-B-Raf (Ser445) Antibody

Polyclonal Antibody for studying B-Raf (Ser446) phosphate. Validated for Western Blotting. Available in 2 sizes. Highly specific and rigorously validated in-house, Phospho-B-Raf (Ser445) Antibody (CST #2696) is ready to ship. Product Usage Information Western Blotting: 1:1000 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting Specificity / Sensitivity Phospho-B-Raf (Ser445) Antibody detects endogenous levels of B-Raf when phosphorylated at serine 445. Human B-Raf residue Ser446 historically has been referenced as Ser445 and is homologous to mouse Ser431. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Polyclonal antibodies are produced by immunizing animals with a phospho synthetic peptide corresponding to residues surrounding serine 445 of human B-Raf. Antibodies are purified by protein A and peptide affinity chromatography. Background 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 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 (Ser446), 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 (Ser365, Ser429, and Thr440) 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). Alternate Names 94 kDa B-raf protein; B-raf; B-Raf proto-oncogene serine/threonine-protein kinase (p94); B-Raf proto-oncogene, serine/threonine kinase; B-Raf serine/threonine-protein; B-RAF1; BRAF; BRAF1; FLJ95109; MGC126806; MGC138284; murine sarcoma viral (v-raf) oncogene homolog B1; NS7; p94; Proto-oncogene B-Raf; RAFB1; Serine/threonine-protein kinase B-raf; v-raf murine sarcoma viral oncogene homolog B; v-Raf murine sarcoma viral oncogene homolog B1 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 86 SOURCE: Rabbit