Immunoprecipitation (IP) of BRAF by using TrueMab monoclonal anti-BRAF antibodies (Negative control: IP without adding anti-BRAF antibody.). For each experiment, 500ul of DDK tagged BRAF overexpression lysates (at 1:5 dilution with HEK293T lysate), 2ug of anti-BRAF antibody and 20ul (0.1mg) of goat anti-mouse conjugated magnetic beads were mixed and incubated overnight. After extensive wash to remove any non-specific binding, the immuno-precipitated products were analyzed with rabbit anti-DDK polyclonal antibody.
HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY BRAF (RC211013, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-BRAF (1:500).
Western blot analysis of extracts (35ug) from 7 cell lines lysates by using anti-BRAF monoclonal antibody (1:500).
Equivalent amounts of cell lysates (10 ug per lane) of wild-type HeLa cells (WT, Cat# LC810HELA) and BRAF-Knockout HeLa cells (KO, Cat# LC835315) were separated by SDS-PAGE and immunoblotted with anti-BRAF monoclonal antibody TA500444 (1:500). Then the blotted membrane was stripped and reprobed with anti-PCNA antibody as a loading control.