WB
Immunoprecipitating IgG and DYKDDDDK Tag (FLAG) in recombinant protein with falg on N-terminal. 25ng of Immunoprecipitated protein incubated with primary antibody (1/1000) for over night at 4℃. For western blotting a IP Detection Reagent(1/1000) was used to confirm successful immunoprecipation.
Lane1/2: Immunoprecipitating IgG in recombinant protein with flag on N-terminal
Lane3: Immunoprecipitating DYKDDDDK Tag (FLAG) in recombinant protein with flag on N-terminal
IP
PCNA was immunoprecipitated from 0.2 mg HeLa cell lysate with PCNA Recombinant Mouse Monoclonal Antibody at 2µg/25µl agarose. Western blot was performed from the immunoprecipitate using PCNA Recombinant Mouse Monoclonal Antibody at 1/10,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: PCNA Recombinant Mouse Monoclonal Antibody IP in HeLa cell lysate
Lane 3: Mouse IgG instead of PCNA Recombinant Mouse Monoclonal Antibody in HeLa cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 24 seconds; ECL: K1801