|Detection of Human/Mouse/Rat B‑Raf by Western Blot. Western blot shows lysates of MDA-MB-468 human breast cancer cell line, Raji human Burkitt's lymphoma cell line, and Ramos human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 µg/mL Mouse Anti-Human/Mouse/Rat B‑Raf Monoclonal Antibody (Catalog # MAB3424) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, Recombinant Human Active Raf‑1 (Y340E Y341E), aa 306-648 (Catalog # 4540-KS) and recombinant human B-Raf (5 ng/lane) were included. A specific band for B-Raf was detected at approximately 95 kDa (as indicated). An unidentified band at 55 kDa was also detected. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.|
The Raf serine/threonine kinases are effectors of Ras that function as MAP3Ks in the ERK phosphorylation cascade. Mammals express three Raf proteins: Raf-1 (C‑Raf); A-Raf; and B-Raf, strongly expressed in fetal brain and adult cerebrum. Mice with a targeted disruption of the B-raf gene die of vascular defects during mid‑gestation. B-raf mutations have been found in two-thirds of malignant melanomas, with the single substitution V599E in the kinase domain the most frequent occurrence.
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