Western blot analysis of extracts (35ug) from 9 different cell lines by using anti-MAPK1 monoclonal antibody.
Equivalent amounts of cell lysates (10 ug per lane) of wild-type Hela cells (WT, Cat# LC810HELA) and MAPK1-Knockout Hela cells (KO, Cat# LC810185) were separated by SDS-PAGE and immunoblotted with anti-MAPK1 monoclonal antibody TA500469. Then the blotted membrane was stripped and reprobed with anti-HSP90AB1 antibody (TA500494) as a loading control (1:200).
Immunoprecipitation of MAPK1 by using TrueMab monoclonal anti-MAPK1 antibody (Negative control: IP without adding anti-MAPK1 antibody). For each experiment, 500ul of DDK tagged MAPK1 overexpression lysates (at 1:5 dilution with HEK293T lysate), 2ug of anti-MAPK1 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 MAPK1 (RC204703, 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-MAPK1. Positive lysates LY408481 (100ug) and LC408481 (20ug) can be purchased separately from OriGene.