[Preparation and characterization of rabbit antibody against human mast cell carboxypeptidase].
OBJECTIVE
To prepare antibody against human mast cell carboxypeptidase (hMC-CP) by using recombinant hMC-CP expressed in E.coli, and to characterize the antibody.


METHODS
hMC-CP was expressed in E.coli with L-Arabinose induction and purified through Ni-NTA column. The purified hMC-CP as immunogen was used to immunize rabbit. The titer and the specificity of the rabbit anti-hMC-CP antibody was analyzed by indirect ELISA and Western blot respectively.


RESULTS
The hMC-CP was successfully expressed and purified. The polyconal anit-hMC-CP antibody was prepared by immunizing rabbit using the purified recombinant protein, The titer of the generated antiserum was 1:6 400 by ELISA. Western blot analysis showed that this antibody could bind with hMC-CP specifically.


CONCLUSIONS
The rabbit anti-hMC-CP antibody with high titer and high specificity has been prepared by using purified recombinant hMC-CP as immunogen, which lays the foundation for further research on detection and function of hMC-CP.
Chen
Zhang-quan
ZQ
Department of Clinical Immunology, Guangdong Medical College, Zhanjiang 524023, China. czqhk@126.com
He
Su-hui
SH
Lu
Tian-tian
TT
He
Tian-wen
TW
He
Shao-heng
SH
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D002268:Carboxypeptidases / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D008407:Mast Cells / Q000201:enzymology*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D013997:Time Factors