Preparation and application of rabbit anti-osteoclastic protein tyrosine phosphatase polyclonal antibody / 吉林大学学报(医学版)

ABSTRACT

Objective: To purify the recombinant protein by using the constructed pET28a-APTP-oc recombinant expression plasmid, and to prepare the rabbit polyclonal antibody against osteoclastic protein tyrosine phosphatase (PTP-oc) and to identify its properties. Methods: The constructed pET28a-APTP-oc recombinant plasmid vyas transformed into E.CO BL21 (DE3), and the isopropyl-ß-D-thiogalactoside (IPTG) vyas used to induce the expression of recombinant protein. The recombinant protein vyas purified by the method of Q fast flovy agarose and Ni-NTA agarose, and the purified recombinant protein vyas used as an antigen to immunize the rabbit to obtain PTP-oc polyclonal antibody. The titer and specificity of the antibody vyere identified by indirect ELISA and Western blotting methods. Results: The recombinant PTP-oc protein vyas successfully purified. The indirect ELISA results shovyed that after immunizing the rabbit vyith the recombinant PTP-oc protein, the polyclonal antibody vyith the titer of 1 : 3 2000 vyas gotten. The Western blotting results shovyed that the polyclonal antibody had high specificity. Conclusion: The PTP-oc recombinant protein is successfully purified, and the polyclonal antibody of PTP-oc is prepared successfully.