CST, 48748SF, B-Raf (E2T8S) Mouse Monoclonal Antibody (BSA and Azide Free)

Monoclonal Antibody for studying B-Raf. Validated for ELISA+. Highly specific and rigorously validated in-house, B-Raf (E2T8S) Mouse Monoclonal Antibody (BSA and Azide Free) (CST #48748) is ready to ship. Product Usage Information This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us . Optimal dilutions/concentrations should be determined by the end user. BSA and Azide Free antibodies are quality control tested by size exclusion chromatography (SEC) to determine antibody integrity. Formulation Supplied in 1X PBS (10 mM Na 2 HPO 4 , 3 mM KCl, 2 mM KH 2 PO 4 , and 140 mM NaCl (pH 7.8)). BSA and Azide Free. Storage Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance. Specificity / Sensitivity B-Raf (E2T8S) Mouse Monoclonal Antibody (BSA and Azide Free) recognizes endogenous levels of total B-Raf protein. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a fragment of human B-Raf protein. 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 Source/Isotype: Mouse IgG2b kappa