JOURNAL OF SHANDONG UNIVERSITY(NATURAL SCIENCE) ›› 2014, Vol. 49 ›› Issue (1): 25-30.doi: 10.6040/j.issn.1671-9352.0.2013.445

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The preparation and purification of Supervillin antibody

ZHANG Tong-tong, QIAN Ji-cheng, ZHU Chang-jun, DONG Zhi-xiong*   

  1. Key Laboratory of Molecular and Cellular Systems Biology, Tianjin Normal University, Tianjin 300387, China
  • Received:2013-09-18 Online:2014-01-20 Published:2014-01-15

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Abstract:

Gelling spot protein Supervillin(SVIL) is a membrane protein molecule located in cells and extracellular matrix interface adhesive spots. To further study the cellular function of SVIL protein,we constructed plasmids of pGEXKG-SVILC302 and pHIS8-SVILC302 to express recombinant protein GST-SVILC302 and HIS-SVILC302 in E.coli BL21-DE3, respectively. The GST-SVILC302 protein was purified by Glutathione affinity chromatography and used to immune New Zealand white rabbits to generate the polyclonal antibody of SVIL. The HIS-SVILC302 protein was crosslinked with Ni-TNA Beads for affinity purification of the antibody. The results of Western Blot and Immunoflurescence experiments demonstrated that the antibody can specific recognize exogenous GFP-SVIL and endogenous SVIL protein. Our results showed that anti-SVIL polyclonal antibody we generated has high specificity and sensitivity enough to study the cellular functions of SVIL protein.

Key words: Supervillin, antibody, purification

CLC Number: 

  • Q78
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