Preparation and identification of rabbit anti-mouse coiled-coil domain containing 189(Ccdc189)polyclonal antibody.
- Author:
Huiping ZHOU
1
;
Fan YANG
1
;
Lu YUAN
1
;
Changmin NIU
1
;
Ying ZHENG
2
Author Information
1. Department of Histology and Embryology, School of Medicine, Yangzhou University, Jiangsu Key Laboratory of Experimental & Translational Non-coding RNA Research, Yangzhou University, Yangzhou 225001, China.
2. Department of Histology and Embryology, School of Medicine, Yangzhou University, Jiangsu Key Laboratory of Experimental & Translational Non-coding RNA Research, Yangzhou University, Yangzhou 225001, China. *Corresponding author, E-mail: yzzkl@163.com.
- Publication Type:Journal Article
- MeSH:
Rabbits;
Male;
Animals;
Mice;
Antibody Specificity;
Antibodies;
Enzyme-Linked Immunosorbent Assay;
Blotting, Western;
Recombinant Proteins;
Escherichia coli/genetics*
- From:
Chinese Journal of Cellular and Molecular Immunology
2023;39(5):456-462
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare a rabbit anti-mouse coiled-coil domain containing 189 (Ccdc189) polyclonal antibody. Methods The pET-28a-Ccdc189 prokaryotic expression plasmid was constructed and transformed into E.coli BL21. IPTG was used to induce the expression of Ccdc189 prokaryotic protein. Adult male New Zealand rabbits were immunized with purified recombinant protein to obtain rabbit anti-mouse Ccdc189 polyclonal antibody. The specificity of the polyclonal antibody was identified by Western blot analysis, indirect ELISA and immunofluorescence histochemical staining. Results The pET-28a-Ccdc189 recombinant plasmid was successfully constructed and the expression of the Ccdc189 recombinant protein was induced. ELISA revealed that the titer of the polyclonal antibody was 1:1 000 000. Western blot and immunofluorescence staining demonstrated that the Ccdc189 polyclonal antibody could specifically identify the Ccdc189 prokaryotic protein and the Ccdc189 protein in adult wild-type mouse testis. Conclusion A polyclonal antibody with high specificity against mouse Ccdc189 was successfully created.
