兔抗破骨细胞蛋白质酪氨酸磷酸酶多克隆抗体的制备和应用

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Preparation and application of rabbit anti-osteoclastic protein tyrosine phosphatase polyclonal antibody

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doi:10.13481/j.1671-587x.20180339

Abstract: Objective: To purify the recombinant protein by using the constructed pET28a-ΔPTP-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-ΔPTP-oc recombinant plasmid was transformed into E.coli BL21(DE3),and the isopropyl-β-D-thiogalactoside (IPTG) was used to induce the expression of recombinant protein.The recombinant protein was purified by the method of Q fast flow agarose and Ni-NTA agarose,and the purified recombinant protein was used as an antigen to immunize the rabbit to obtain PTP-oc polyclonal antibody.The titer and specificity of the antibody were identified by indirect ELISA and Western blotting methods. Results: The recombinant PTP-oc protein was successfully purified.The indirect ELISA results showed that after immunizing the rabbit with the recombinant PTP-oc protein,the polyclonal antibody with the titer of 1:3 2000 was gotten.The Western blotting results showed 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.

Key words: titer, osteoclastic protein-tyrosine phosphatase, polyclonal antibody, osteoclast, identification

CLC Number:

Q814

LIU Shaowei, JIANG Huan, WANG Xiaolong, LI Menghong, CHEN Yu, TANG Zhongyuan, HU Min. Preparation and application of rabbit anti-osteoclastic protein tyrosine phosphatase polyclonal antibody[J].Journal of Jilin University Medicine Edition, 2018, 44(03): 661-666.

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