[Preparation and characterization of rabbit antibody against gastric cancer-related protein GCRG213].
OBJECTIVE
To prepare the rabbit antibody against gastric cancer-related protein GCRG213.


METHODS
The thioredoxin/GCRG213 fusion protein was expressed in E. coli. The rabbit antibody against GCRG213 was obtained by immunizing a rabbit with the purified GCRG213 protein. The titer and specificity of the antibody was determined by ELISA and Western-blot, respectively.


RESULTS
The thioredoxin/GCRG213 fusion protein with relative molecular mass (Mr) of 29,400 was overexpressed in E. coli. The purity of expressed product directly purified from a denaturing polyacrylamide gel was about 100%. The rabbit antibody against GCRG213 was obtained. The ELISA titer of antiserum against GCRG213 was about 1:256,000. Western blot analysis showed that the antiserum could bind to the expressed fusion protein specifically.


CONCLUSIONS
The rabbit antibody against GCRG213 has been successfully prepared, which lays the foundation for further studying the biological function and the possible role of the GCRG213 in the development of gastric carcinoma.
Wu
Yin-qiao
YQ
Department of Gastroenterology, South Building General Hospital of PLA, Beijing 100853, China. wuyq56@hotmail.com
Wang
Meng-wei
MW
Wu
Ben-yan
BY
Wang
Gang-shi
GS
You
Wei-di
WD
Wang
Wei-hua
WH
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D004720:Endonucleases / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D007106:Immune Sera / Q000096:biosynthesis* / Q000276:immunology; D008297:Male; D036361:Peptide Hormones; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013879:Thioredoxins / Q000096:biosynthesis / Q000235:genetics
D000906:Antibodies; C477000:GKN1 protein, human; D007106:Immune Sera; D036361:Peptide Hormones; D011993:Recombinant Fusion Proteins; D013879:Thioredoxins; D004720:Endonucleases