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.
