2.5. Co-Immunoprecipitation Assay

Proteins from rat hippocampal tissues were incubated on a wheel overnight at 4 °C in RIA buffer 1× (200 mM NaCl, 10 mM EDTA, 10 mM Na₂HPO₄, 0.5% NP-40) plus 0.1% SDS and primary antibodies. The day after, magnetic beads A (Bio-Rad #1614013) were washed in PBS, resuspended in RIA 1×, added to the samples, and incubated at RT on a wheel for 1 h. The beads were then sedimented and the supernatant was discarded. The beads were washed three times with RIA 1× + 0.1% SDS mixed with loading buffer (3×), and boiled for 10 min at 96 °C for WB procedures.