Mouse Foxp3 protein expression and polyclonal antibody.doc

 PAGE \* MERGEFORMAT 14 Mouse Foxp3 protein expression and polyclonal antibody Of: Bao Junfeng, Wang Shengjun, Ma Jie, David Zweig, Yang prophet, Mao Zhaoming, with good, Shaoqi Xiang Xu of the River [Abstract] Objective: In vitro primary mouse Foxp3 protein expression, and with its immune rabbit polyclonal antibody. Methods: Foxp3 pMD 18T amplified from plasmid full-length mouse Foxp3 gene, was cloned into prokaryotic expression vector pET 32a, into E.coli BL 21, IPTG induced expression of the target protein, Ni2 + NTA and Western blot identification of purified protein, purified protein rabbit polyclonal antibody, ELISA antibody titer assay, Western blotting antibody specificity. Results: successfully constructed Foxp3 pET 32a prokaryotic expression vector, expressed and purified protein, ELISA assay titer 1:20 000, Western blotting showed that antibodies specific for mouse Foxp3 protein. Conclusion: The expression of murine Foxp3 protein and polyclonal antibodies. [Keywords:] Foxp3, expression, polyclonal antibody, mouse [Abstract] Objective: To prepare rabbit polyclonal antibody against mouse Foxp3 protein by prokaryotic expression of mouse Foxp3 protein in vitro. Methods: Foxp3 gene was amplified by PCR from the full length plasmid Foxp3 pMD 18T and subcloned into the pET 32a vector; the recombinant plasmid was transformed into E.coli BL 21 and the expression of recombinant protein was induced by IPTG. after purified with Ni2 + NTA column, identification of the recombinant protein by Western blot was performed; the purified protein was used as antigen to immunize rabbits, the titer and the specificity of the rabbit anti Foxp3 antibody were detected by ELISA and Western blot, respectively. Results: The prokaryotic expression vector for Foxp3 pET 32a was constructed successfully; mouse Foxp3 protein had been expressed and purified; rabbit polyclonal antibody against mouse Foxp3 protein was prepared, the titer of the antibody was 1:20 000 by ELISA