ELISA help needed

Martin K. Oaks moaks at execpc.com
Mon Oct 14 12:19:35 EST 1996


I am having trouble with high "background" levels in a sandwich elisa 
assay. My capture antibody is a mouse monoclone, my test substance is 
human serum, the second ab is a humanized IgG, the reporter ab is 
peroxidase conjugated anti-human IgG. I get high levels of background 
regardless of whether the capture ab I use is relevant or not. In other 
words, ANY mouse ab appears to be reactive. I get no activity without 
mouse capture ab and/or human serum. In addition, my reporter ab does not 
react with the capture ab. It would appear that my test sera have 
anti-mouse antibodies (HAMA) although I have no reason to believe that 
these people have been exposed to mouse Igs. Should I absorb these test 
sera (with solid phase mouse Igs) or is there a more straightforward 
approach?

reply here or email me: Moaks at execpc.com

Thanks.



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