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[Preparation and application of anti-serum against mouse visfatin]
1Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China. mxh06@mails.tsinghua.edu.cn
Aim:
To prepare polyclonal anti-serum against mouse visfatin and explore the expression of visfatin in different mouse tissue.
Methods:
A fragment of gene coding the full length of visfatin was amplified and cloned into the prokaryotic expression vector pET28a. Then the recombined protein was expressed in E.coli(BL21) and purified by Ni-NTA HIS*BIND RESIN. After the indentification of Western blot, the recombined protein was used to immune New Zealand rabbit. With the purified anti-serum, we characterized the expression of visfatin in different mouse tissues.
Results:
According to the results of SDS-PAGE and Western blot, the expression of recombined prokaryotic expression vector pET28a-visfatin could be induced by IPTG in E.coli. The titer of the anti-serum was above 1:25 600.
Conclusion:
The results for the identification of vifatin among mouse tissues show that visfatin exists in all the detected tissues, with the expression in spleen and adipose tissue higher than the others.
