Expression of Catalytic Domain of Protein Tyrosine Phosphatase 1B and Preparation of Its Polyclonal Antibody

摘要

This study focuses on the expression of human protein tyrosine phosphatase 1B(PTP1B) catalytic domain(ΔPTP1B) and preparation of polyclonal antibody against ΔPTP1B. ΔPTP1B gene was PCR amplified with the cDNA of human PTP1B as the template, and cloned into the pT7 expression vector. The recombinant pT7-ΔPTP1B was expressed in E. coli Rosetta(DE3) host cells and purified. The antiserum was prepared by immunizing rabbit with purified recombinant ΔPTP1B. The polyclonal antibody against ΔPTP1B was purified by PVDF immobilized antigen affi-nity chromatography. ΔPTP1B was correctly cloned, expressed, and purified as confirmed by PCR, DNA sequence analysis, SDS-PAGE western blotting, and HPLC. The titer and sensitivity of the antibody were 1∶2500(volume ratio) and 0.1 ng, respectively. This study provides an important basis for further studying the biological function of PTP1B and its relationship with human diseases.