Brain or cell lysates were pre-cleared with protein A-Sepharose beads (YEASEN Biotech, Cat#36403ES25). After washing, lysates were incubated with primary antibody for 8 hr at 4 °C. After incubation, complexes were precipitated with 10 μl of protein A-Sepharose beads with gentle agitation at 4 °C for 2 h; nonspecific IgG was used as a negative control. Beads were washed, and the immunoprecipitated protein complex was loaded onto SDS-PAGE gels and processed for western blotting.
