Preparations of the specific antibodies against exon 2 and exon 3 of human tau protein
10.3760/cma.j.issn.1003-9279.2009.02.024
- VernacularTitle:人tau蛋白外显子2和外显子3特异性抗体的制备
- Author:
Cao CHEN
1
;
Gui-Rong WANG
;
Qi SHI
;
Bao-Yun ZHANG
;
Guo-Yong MEI
;
Yuan LI
;
Xin WANG
;
Rui-Min ZHOU
;
Jun HAN
;
Yu-Jun ZHAO
;
Xiao-Ping DONG
Author Information
1. 沈阳农业大学
- Keywords:
Proteins;
Exons;
Antibodies,viral;
Prion diseases
- From:
Chinese Journal of Experimental and Clinical Virology
2009;23(2):146-148
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare the specific antibodies against exon 2 and exon 3 of human tan protein. Methods Sequences encoding exon 2 and exon 3 of human tau protein were amplified from human peripheral blood DNA and cloned into a prokaryotic expression vector pGEX-2T. Fusion proteins GST-E2 and GST-E3 were expressed and purified from E. coli system. The antisera were elicited by immunization of the purified fusion proteins to rabbits and mice. The specific antibodies were purified by Protein G/A and CNBr-aetivated sepharose 4B coupled with GST protein. The specificity and sensitivity of the purified antibodies were evaluated by Western blotting and ELISA.Results Recombinant fusion proteins GST-E2 and GST-E3 were expressed and purified from E. coli, which showed Mr. 29 x 103 . Various antisera were collected from the immunized experimental animals. Reliable immunoreactive specificity and titers of the purified antibodies against exon 2 and exon 3 of human tau protein were confirmed by Western blotting and ELISA after serial purification processes. Conclusion Four specific antibodies against exon 2 and exon 3 of human tau protein have been successfully prepared, which provides basis for analyzing the role of tau in neurodegenerative diseases.
