ELISA help needed

Bryan Kiehl b3748 at cts.com
Tue Oct 15 20:32:23 EST 1996


I am not sure this will work in your case, but in diagnostic kits, you
may add various "blocking" reagents such as goat or bovine sera to the
the specimen and conjugate diluents. To be successful, you may need
levels that vary between 5-30%.  This is much like absorbtion, but
alot easier.

On Mon, 14 Oct 1996 10:19:35 -0700, "Martin K. Oaks"
<moaks at execpc.com> wrote:

>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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