[Prokaryotic expression of endostatin and preparation of polyclonal antibody].
OBJECTIVE
The aim of this study was to express and purify human endostatin and to prepare polyclonal antibody of mouse anti-human endostatin.


METHODS
The cDNA of endostatin was amplified by PCR, then recombined into prokaryotic expression vector and transformed into Escherichia coli BL21 for expression; the mice were immunized with purified products.


RESULTS
Prokaryotic expression vector pQE-30 of human endostatin was successfully constructed; the expression product was gained after pQE-30 was transferred into BL21. After purified by Ni affinity chromatography, the product was identified to be a single component by SDS-PAGE. Western blot analysis showed that high titer mouse anti-human endostatin polyclonal antibody was successfully prepared.


CONCLUSIONS
Highly purified expression product and prepared polyclonal antibody provide the necessary material for further study.
Zhang
Yi-ming
YM
Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing 210029, P. R. China. zym6677@sina.com.cn
Wang
Ruo-ning
RN
Li
Yan-li
YL
Gu
Dong-min
DM
Cheng
Zhi-xiang
ZX
Xiong
Jun
J
De
Wei
W
Chen
Bing-ying
BY
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D003094:Collagen / Q000235:genetics* / Q000276:immunology; D043169:Endostatins; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D043169:Endostatins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D003094:Collagen