Co immunoprecipitation co ip kit

Co-IP assay was performed using the Co-Immunoprecipitation (Co-IP) Kit (Thermo Scientific) according to the manufacturer’s instructions. Purified anti-Tmod antibodies or anti-Pns10 antibodies were added to the resin for immobilization for 2 h at room temperature. Then RDV-infected or mock-infected cultured cells were lysed, and the supernatant was incubated with the antibody-immobilized resin for 1 h at 4 °C. The Co-IP fraction was eluted followed by SDS-PAGE and Western blotting analysis using anti-Pns10 antibodies, or anti-Tmod as primary antibodies and anti-mouse IgG-Alkaline Phosphatase antibody (Sigma) as secondary antibodies.

Co-immunoprecipitation assay was performed with a Co-Immunoprecipitation (Co-IP) Kit (Thermo Fisher Scientific Inc, Rockford, USA) according to the manufacturer's instructions. Briefly, 75 µg of affinity-purified antibodies [Sp1 (polyclonal rabbit IgG, protein A purified), HDAC1 (monoclonal mouse IgG, immunogen affinity purified) and HDAC4 (link) (monoclonal rabbit IgG, immunogen affinity purified), Cell Signaling Technology)] were coupled into the spin columns. Cell lysate was carefully prepared and pre-cleared using the control agarose resin. Next, the cell lysate was co-immunoprecipitated and eluted. Finally, samples were evaluated by western blot analysis.