Expression and purification of GST-CML28 fusion protein and preparation of its polyclonal antibody.
- Author:
Xia MAO
1
;
Bing ZHANG
;
Xue-Ling BAI
;
Long-Long LIU
;
Dong-Hua ZHANG
Author Information
1. Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies;
immunology;
isolation & purification;
metabolism;
Antigens, Neoplasm;
biosynthesis;
immunology;
isolation & purification;
Cells, Cultured;
Escherichia coli;
metabolism;
Exosome Multienzyme Ribonuclease Complex;
biosynthesis;
immunology;
isolation & purification;
Genetic Vectors;
Glutathione Transferase;
biosynthesis;
isolation & purification;
Human Umbilical Vein Endothelial Cells;
cytology;
Humans;
RNA-Binding Proteins;
biosynthesis;
immunology;
isolation & purification;
Rabbits;
Recombinant Fusion Proteins;
biosynthesis;
immunology;
isolation & purification
- From:
Journal of Experimental Hematology
2012;20(6):1302-1306
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the expression of GST-CML28 in Escherichia Coli and to prepare its antibody. The constructed recombinant expression vectors CML28-pGEX-3X were transformed into Escherichia Coli BL21 under IPTG induction. The protein was abstracted from the transformers, and purified by a GSTrap FF column. The rabbits were immunized by the purified fusion protein to produce serum with anti-CML28 antibody. The serum was purified by chromatographic column stuffed with glutathione Sephamse 4B to get the antibody. The specific antibody against CML28 was further identified by ELISA, Western blot, immunohistochemistry and quantum dot luminescence. The results indicated that GST-CML28 fusion protein was expressed in Escherichia coli and its specific polyclonal antibody was obtained. It is concluded that the anti-CML28 polyclonal antibodies with high titer and specificity are successfully prepared. These antibodies provide an useful experimental tool to profoundly research the physiological significance and biological function of the CML28 gene.
