Co-immunoprecipitation (Co-IP) assay
Co-IP assay was performed using the BeaverBeads protein A/G
immunoprecipitation Kit (BEAVER Biomedical Engineering) per
manufacture’s protocol. Briefly, both the beads and antibodies were
pre-treated with binding buffer supplied in the kit, then the extracted
total protein was mixed with the pre-treated antibodies and beads, and
incubated overnight at 4 °C with gentle shaking. After five times’
washing, the beads were magnetic separated and resuspended in elution
buffer containing coomassie brilliant blue. The precipitates were boiled
for 5 min and applied to SDS-PAGE for further analysis.