1. [The expression and antibody preparation of S100A10 protein].
- Author
-
Xin D, Zou X, Zuo M, and Liu C
- Subjects
- Animals, Annexin A2 immunology, Annexin A2 isolation & purification, Annexin A2 physiology, Antibody Specificity, Plasmids, Rabbits, S100 Proteins immunology, S100 Proteins isolation & purification, Annexin A2 genetics, Antibodies immunology, Recombinant Proteins biosynthesis, S100 Proteins genetics
- Abstract
Objective: To prepare S100A10 protein and its specific polyclonal antibody., Methods: The full-length gene fragment of S100A10 was amplified by PCR, and then cloned into pET28a(+) prokaryotic expression vector. After transformation, the vector was induced to express the recombinant (S100A10)(2) protein by IPTG in E.coli BL21 (DE3). The recombinant (S100A10)(2) was then purified by Ni-NTA resin. (S100A10)(2)-specific polyclonal antibody was prepared using the purified recombinant (S100A10)(2) protein as antigen to inoculate rabbit intradermally. The title and specificity of the polyclonal antibody were determined by ELISA and Western blotting., Results: The study successfully constructed the prokaryotic recombinant expression vector pET28a(+)-(S100A10)(2), and obtained the purified recombinant (S100A10)(2) protein and polyclonal antibody with high titer and specificity., Conclusion: The prokaryotic expression and purification system for S100A10 has been established and polyclonal antibody of (S100A10)(2) been prepared, which provides helpful fools for further researches on S100A10.
- Published
- 2014