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.