Overexpression,purification and polyclonal antibody preparation of recombinant protein Fpr3

In order to get insights into the structural and molecular interaction mechanisms between PPIase and PP1, the recombinant plasmid pGEX-5X-1-Fpr3 was constructed and overexpressed in E. coli as a soluble protein. The fusion protein was purified by GSTrap affinity chromatography and the GST tag was removed by the method of on-column cleavage with Factor Xa. The recombinant GST-Fpr3 and Fpr3 was assayed and identified by SDS-PAGE and Western blot, respectively. The recombinant GST-Fpr3 was used to immunize rabbits for preparing polyclonal antibody. The polyclonal antibody with high titer and high specificity against the GST-Fpr3 has been successfully prepared.