Preparation and identification of the polyclonal antibody against mouse IL-23p19].

To prepare the polyclonal antibody against mouse interleukine-23 p19 (IL-23p19). The murine full-length IL-23p19 gene was subcloned into pET16 expression vector to construct the recombinant plasmid pET-16b-IL-23p19. The plasmid was transformed into E. coli BL 21 (DE3) and IL-23p19 protein expression was induced by IPTG and identified with SDS-PAGE analysis and Western blotting. After purified by Ni(+) affinity column chromatography, the IL-23p19 protein was used as the antigen to immunize New Zealand rabbit to prepare the antiserum. The polyclonal antibody against the mouse IL-23p19 was isolated from antiserum by affinity chromatography. Antibody titer was detected by ELISA. Antibody specificity was evaluated by Western blotting. The pET-16b-IL-23p19 recombinant plasmid was successfully constructed and the IL-23p19 protein was effectively expressed in E. coli BL 21 (DE3). The antibody was successfully prepared by immunizing New Zealand rabbit with the IL-23p19 protein four times. ELISA showed that the titer of the anti-mouse IL-23p19 polyclonal antibody was about 1:256,000. Western blotting confirmed that anti-mouse IL-23p19 polyclonal antibody could specifically recognize the IL-23p19 protein. We have successfully prepared the anti-IL-23p19 polyclonal antibody with the high titer and specificity.

Citation

Yingli Men, Xiaolong Wang, Cong Ding, Xiaodong Wang, Zhenyu Ji, Ting Wang, Qiaozhen Kang, Xin Liu. Preparation and identification of the polyclonal antibody against mouse IL-23p19]. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology. 2016 May;32(5):688-92

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PMID: 27126950

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