Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), C2C12 (Lane 3), Raji (Lane 4), Ramos (Lane 5), U-87 MG (Lane 6), T98G (Lane 7), U-2 OS (Lane 8), HT-29 (Lane 9), HCT116 (Lane 10) and NIH/3T3 (Lane 11). The blot was probed with Anti-B-Raf Recombinant Rabbit Polyclonal Antibody (Product # 711219, 1 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). 84 kDa band corresponding to B-Raf was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).