title: [Preparation and identification of the polyclonal antibody against mouse IL-23p19].
abstract: OBJECTIVE
  To prepare the polyclonal antibody against mouse interleukine-23 p19 (IL-23p19).
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  We have successfully prepared the anti-IL-23p19 polyclonal antibody with the high titer and specificity.
authors.affiliation: Laboratory of Molecular Immunology, School of life sciences, Zhengzhou University, Zhengzhou 450000, China.
authors.forename: Yingli
authors.initials: Y
authors.lastname: Men
authors.affiliation: Laboratory of Molecular Immunology, School of life sciences, Zhengzhou University, Zhengzhou 450000, China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Wang
authors.affiliation: Laboratory of Molecular Immunology, School of life sciences, Zhengzhou University, Zhengzhou 450000, China.
authors.forename: Cong
authors.initials: C
authors.lastname: Ding
authors.affiliation: Laboratory of Molecular Immunology, School of life sciences, Zhengzhou University, Zhengzhou 450000, China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Medicine, Zhengzhou University, Zhengzhou 450000, China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Ji
authors.affiliation: Laboratory of Molecular Immunology, School of life sciences, Zhengzhou University, Zhengzhou 450000, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: Laboratory of Molecular Immunology, School of life sciences, Zhengzhou University, Zhengzhou 450000, China. *Corresponding authors, E-mail: qzkang@zzu.edu.cn.
authors.forename: Qiaozhen
authors.initials: Q
authors.lastname: Kang
authors.affiliation: Laboratory of Molecular Immunology, School of life sciences, Zhengzhou University, Zhengzhou 450000, China. *Corresponding authors, E-mail: liux@zzu.edu.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D053760:Interleukin-23 Subunit p19 / Q000235:genetics / Q000276:immunology*; D008297:Male; D051379:Mice; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000906:Antibodies; D053760:Interleukin-23 Subunit p19; D011994:Recombinant Proteins