利用免疫共沉淀技术验证SET与eEF1A1在人肝细胞内的相互作用

目的]探讨SET与eEF1A1在人肝细胞内是否存在相互作用。方法]以人L-02肝细胞为材料,在非变性条件下提取总蛋白,将鼠抗人SET、兔抗人eEF1A1抗体分别加到总蛋白提取物中,用偶联了琼脂糖珠的Protein A/G分离抗原抗体复合物,最后分别用兔抗人eEF1A1、鼠抗人SET抗体进行Western blotting检测。结果]在用SET抗体免疫共沉淀下来的蛋白复合物中检测到eEF1A1,同时在用eEF1A1抗体免疫共沉淀下来的蛋白复合物中也检测到SET。结论]SET与eEF1A1在人肝细胞内存在相互作用。

Verification of the Interaction between SET and eEF1A1 in Human Liver Cells by Co-immunoprecipitation

Objective] The aim was to investigate the possible interaction between SET and eEF1A1 in human liver cells.Method] First,the total proteins of human L-02 liver cells were extracted under non-denaturing conditions;then,mouse anti-human SET and rabbit anti-human eEF1A1 antibodies were used to perform the co-immunoprecipitation respectively;subsequently,the immunoprecipitations was correspondingly detected with rabbit anti-human eEF1A1 and mouse anti-human SET antibodies by Western Blot.Result] EEF1A1 was detected in the protein complex from the immunoprecipitations by using anti-SET antibody,and SET also was detected in the immunoprecipitations by using anti-eEF1A1 antibody.Conclusion] The interaction between SET and eEF1A1 in human liver cells was conformed.