Production and application of rabbit polyclonal antibody against mouse testis expressed 38 (TEX38).
- Author:
Ling YANG
1
;
Lu YUAN
1
;
Fan YANG
1
;
Tingting GE
1
;
Wenhua XU
1
;
Linwei XU
1
;
Changmin NIU
1
;
Ying ZHENG
2
Author Information
1. Institute of Obstetrics, Gynecology and Reproduction, Department of Histology and Embryology, School of Medicine, Yangzhou University, Yangzhou 225001, China.
2. Institute of Obstetrics, Gynecology and Reproduction, Department of Histology and Embryology, School of Medicine, Yangzhou University, Yangzhou 225001, China. *Corresponding author, E-mail: yzzkl@163.com.
- Publication Type:Journal Article
- MeSH:
Male;
Rabbits;
Animals;
Mice;
Testis;
Antibodies;
Enzyme-Linked Immunosorbent Assay;
Immunization;
Spermatozoa;
Escherichia coli
- From:
Chinese Journal of Cellular and Molecular Immunology
2023;39(10):933-939
- CountryChina
- Language:Chinese
-
Abstract:
Objective To produce rabbit polyclonal antibody against mouse testis expressed 38 (TEX38). Methods Full-length open reading frame sequence of TEX38 was amplified and inserted into the pET-30a-(+) vector to construct pET-30a-TEX38 prokaryotic plasmid. The recombinant plasmid was transformed into E.coli BL21, and expression was induced with isopropyl β-D-thiogalactopyranoside (IPTG). New Zealand white rabbits were immunized with TEX38 protein after purification and denaturation, then TEX38 polyclonal antibodies were collected from rabbit serum samples. ELISA was performed to detect the antibody titer. Western blot and immunofluorescence staining were performed to determine the specificity of TEX38 polyclonal antibodies. Results The pET-30a-TEX38 recombinant plasmid was constructed, and TEX38 prokaryotic protein was expressed and purified successfully. After immunization, the titer of TEX38 antibody reached 1:1 000 000. Western blot analysis and immunofluorescence staining showed that TEX38 was localized in the mouse spermatogenic cells and sperms with a good specificity. Conclusion The rabbit polyclonal antibody against mouse TEX38 is successfully produced, and the expression of TEX38 in mouse spermatogenic cells and sperms is validated.
