Preparation and identification of anti-human epididymal sperm protein P34H monoclonal antibody.
- Author:
Xin-yi XIA
1
;
Yong-ming WU
;
Lian-jun PAN
;
Yun GAO
;
Bao-fang JIN
;
Yu-feng HUANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; analysis; biosynthesis; Blotting, Western; Cell Line, Tumor; Enzyme-Linked Immunosorbent Assay; Female; Humans; Hybridomas; secretion; Mice; Mice, Inbred BALB C; Recombinant Proteins; immunology; Sensitivity and Specificity; Sugar Alcohol Dehydrogenases; immunology
- From: National Journal of Andrology 2006;12(11):1004-1006
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare and identify anti-human epididymal sperm protein P34H monoclonal antibody (McAb).
METHODSThe previously purified recombinant P34H protein was used as an antigen to immunize BALB/c mouse. Cell strains secreting anti-P34H McAb were established by hybridoma technique and then ascitic fluid-type McAb prepared. The sensitivity and specificity of McAb were detected by ELISA and Western blot, respectively.
RESULTSOne strain (2C4) of IgG1 Kappa anti-P34H McAb was harvested. The titer detected by ELISA technique was 1:10(3) - 1:10(5). Western blot of healthy human sperm samples and purified recombinant P34H antigen probed with the prepared McAb were immunoreactive at 34,000 and 27,000, respectively.
CONCLUSIONAnti-P34H McAb has been prepared successfully by the above methods, which may provide a powerful tool for exploring the relationship between P34H and male fertility.
