2.13. Co-immunoprecipitation (co-IP)

Kidney tissues were lysed with IP lysis buffer containing protease inhibitor cocktails and the total protein concentration was assessed with BCA assay kit. The lysate supernatant was incubated with 3.675 μg NCOA4 antibody for 3 h at 4 °C and the protein A/G agarose mixture was added to captured the immunocomplexes. Then the immunocomplexes were washed 5 times with precooled IP lysis buffer, and samples were boiled in SDS-PAGE loading buffer and subjected to Western blot.