Ha mouse monoclonal antibody

Co-IP was done using Active Motif Nuclear Complex Co-IP Kit (54001) with IP-low buffer and no supplement. Protein G beads (Dynabeads, ThermoFisher, 10004D) were used for IP. All steps followed the standard protocol. IP samples were separated in 6% SDS-PAGE, the proteins were transferred to a PVDF membrane for 15h at 15V in TRIS-Glycine buffer. For the detection of the proteins of interest, membranes were blocked in a non-mammalian blocking buffer in TTBS (LiCOR Biosciences, #927-60001) for 1h at room temperature. The membranes were incubated with primary antibodies in blocking buffer overnight at 4°C, HA mouse monoclonal antibody (1:1000, Thermo Fisher Scientific, #MA1-12429), V5 mouse monoclonal antibody (1:1000, Thermo Fisher Scientific, R960-25). Then, the membranes were incubated with mouse secondary antibody (LiCOR Biosciences, #925-32210) in TBS for 1h in room temperature. The membrane images were captured in Odyssey Clx (LiCOR Biosciences).