Expression and purification of PPM1A-His fusion protein and preparation of PPM1A polyclonal antibody

Objective To express and purify PPM1A-His fusion protein and prepare PPM1A polyclonal antibody. Methods pBEX1-PPM1A-His was expressed in E.coli BL21 (DE3) pLysS under induction of IPTG. The recombinant protein was purified using Ni-NTA spin column. A polyclonal antibody was developed by immunizing Balb/c mice with the purified fusion protein,and the sensitivity and specificity of the antibody were tested by ELISA,immunohistochemisity,and immunofluorescence technique. Results A high titer (1∶100 000),high sensitivity,and high specificity polyclonal antibody which could recognize human PPM1A in liver tissues and hepatocellular carcinoma was prepared successfully. Conclusion PPM1A mouse polyclonal antibody possesses high sensitive and specificity,which can be used for further study of PPM1A in HCC or other diseases.