HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY USP13 (RC202190, 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-USP13. Positive lysates LY401292 (100ug) and LC401292 (20ug) can be purchased separately from OriGene.
Immunoprecipitation (IP) of USP13 by using TrueMab monoclonal anti-USP13 antibodies (Negative control: IP without adding anti-USP13 antibody.). For each experiment, 500ul of DDK tagged USP13 overexpression lysates (at 1:5 dilution with HEK293T lysate), 2ug of anti-USP13 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.