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[Prokaryotic expression of endostatin and preparation of polyclonal antibody]
Yi-ming Zhang1, Ruo-ning Wang, Yan-li Li
1Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing 210029, P. R. China. zym6677@sina.com.cn
Background & 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.
Conclusion:
Highly purified expression product and prepared polyclonal antibody provide the necessary material for further study.