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).
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.
Standard curve for ELISA analysis with BRAF recombinant protein (dilution range from 0.8ng/ml to 600ng/ml) using BRAF Capture Antibody (Cat# TA500429/TA500462) at 5ug/ml and HRP conjugated BRAF Detection mAb (Cat# TA500438) at 0.03ug/ml.