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[Preparation and identification of anti-human epididymal sperm protein P34H monoclonal antibody]. | LitMetric

[Preparation and identification of anti-human epididymal sperm protein P34H monoclonal antibody].

Zhonghua Nan Ke Xue

Laboratory of Reproduction & Genetics, School of Clinical Medicine, Nanjing University Medical College, Nanjing General Hospital of Nanjing Command, PLA, Nanjing, Jiangsu 210002, China.

Published: November 2006

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.

Conclusion: 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.

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