Preparation and identification of polyclonal antibody against CMTM1-v17/CKLFSF-v17
- VernacularTitle:CMTM1-v17/CKLFSF1-v17多克隆抗体的制备、亲合纯化与鉴定
- Author:
Jing WANG
;
Li WANG
;
Hui FAN
;
Yingmei ZHANG
;
Lu WANG
- Publication Type:Journal Article
- Keywords:
CMTM1-v17;
Gene recombination;
Fusion expression;
Polyclonal antibody
- From:
Chinese Journal of Immunology
1999;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To analyze a novel antigenic determinant of Homo sapiens CMTM1-v17 protein and prepare polyclonal antibody.Methods:By means of DNA recombination method, the C-terminal(The coding sequences of 118-149 residues) of CMTM1-v17 gene was inserted into pGEX-4T-2 vector. The recombinant plasmid was transformed into E.coli XL-1 Blue to express the fusion protein of GST/CMTM1-v17. The GST tag of the purified fusion protein was removed by enzymatic cleavage with thrombin. The polyclonal anti-CMTM1-v17 antibody were prepared by immunizing rabbits with the purified protein. Highly-specific, polyclonal anti-CMTM1-v17 antibodies was purified by affinity chromatographic column matrix coupled with antigen. The specificity of the anti-CMTM1-v17 antibody was identified by Western blot and immunohistochemistry.Results:The polyclonal antibody showed specific response to CMTM1-v17 by Western blot analysis and immunohistochemistry. These results suggested that the polyclonal antibody can be used for further studies of CMTM1-v17 function.Conclusion:A polyclonal anti-CMTM1-v17 antibody has been synthesized successfully.
