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Published on: July 1, 2013
[Preparation and characterization of rabbit antibody against human selenoprotein P]
Qing Fang1, Ming-cheng Zhang, Yuan-yuan Zha
1National Laboratory of Molecular Oncology, Cancer Institute, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100021, China. qingfang71@yahoo.com
Aim:
To prepare rabbit antibody against human selenoprotein P (HSelP) by using HSelP expressed in the prokaryotic expression system and identify its specificity.
Methods:
HSelP gene fragment was expressed in E.Coli via IPTG induction and purified through DEAE and Ni-NTA columns sequentially. The purified HSelP as immunogen was used to immunize rabbit. The specificity of rabbit anti-HSelP antibody was analyzed by Western blot.
Results:
HSelP protein had been expressed and purified successfully. The polyclonal anti-HSelP antibody could specifically recognize natural HSelP from human serum.
Conclusion:
Polyclonal anti-HSelP antibody with high specificity and purity has been prepared by using purified HSelP as immunogen, which lays the foundation for further research on the function and distribution of HSelP, as well as developing a simple method for detecting HSelP.
