Severity: Warning
Message: file_get_contents(https://...@pubfacts.com&api_key=b8daa3ad693db53b1410957c26c9a51b4908&a=1): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests
Filename: helpers/my_audit_helper.php
Line Number: 176
Backtrace:
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 176
Function: file_get_contents
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 250
Function: simplexml_load_file_from_url
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 1034
Function: getPubMedXML
File: /var/www/html/application/helpers/my_audit_helper.php
Line: 3152
Function: GetPubMedArticleOutput_2016
File: /var/www/html/application/controllers/Detail.php
Line: 575
Function: pubMedSearch_Global
File: /var/www/html/application/controllers/Detail.php
Line: 489
Function: pubMedGetRelatedKeyword
File: /var/www/html/index.php
Line: 316
Function: require_once
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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