[Preparation and identification of anti-human epididymal sperm protein P34H monoclonal antibody].
OBJECTIVE
To prepare and identify anti-human epididymal sperm protein P34H monoclonal antibody (McAb).


METHODS
The 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.


RESULTS
One 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.


CONCLUSIONS
Anti-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.
Xia
Xin-yi
XY
Laboratory of Reproduction & Genetics, School of Clinical Medicine, Nanjing University Medical College, Nanjing General Hospital of Nanjing Command, PLA, Nanjing, Jiangsu 210002, China. xiaxy@androl.cn
Wu
Yong-ming
YM
Pan
Lian-jun
LJ
Gao
Yun
Y
Jin
Bao-fang
BF
Huang
Yu-feng
YF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D013401:Sugar Alcohol Dehydrogenases; C023300:L-xylulose reductase