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

This study focuses on the expression of human protein tyrosine phosphatase 1B (PTP1B) catalytic domain (Delta PTP1B) and preparation of polyclonal antibody against Delta PTP1B. Delta PTP1B gene was PCR amplified with the cDNA of human PTP1B as the template, and cloned into the p17 expression vector. The recombinant pT7-Delta PTP1B was expressed in E. coli Rosetta(DE3) host cells and purified. The antiserum was prepared by immunizing rabbit with purified recombinant Delta PTP1B. The polyclonal antibody against Delta PTP1B was purified by PVDF immobilized antigen affinity chromatography. Delta PTP1B 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.