IgM competitive capture ELISA

James Drummond drummond at ncifcrf.gov
Tue Oct 24 13:17:20 EST 1995


In article <46heuk$itd at stork.qut.edu.au> d.beasley at qut.edu.au (David Beasley) writes:
>
>I posted an article here a while ago asking for some help with regard to
>some IgM capture ELISAs I'm doing.  
>
>I have been trying capture ELISAs in 96-well plates from several 
>manufacturers, and it seems that the majority of the antibodies are 
>coating ok but cannot bind the antigen when it is added to the plate (maybe
>because the antibodies can't alter their conformation very well?).  So
>now I am looking for alternatives.
>
>At the moment I am setting up to try blotting the antibodies onto nitrocellulose
>and then do a capture ELISA on the membrane, but I was wondering whether
>anyone has any other suggestions as to how I could immobilise these antibodies.
>
>Thanks,
>
>David B.

I have a few questions.  

1.  why are you using IgM as the capture antibody.  Is this necessary
for what you are doing.  IgM affinities to antigen are generally low
relative to IgG.

2.  What are you trying to capture?

3.  What concentration is the antigen in question?  

4.  What type of plate(s) have you tried to use?

James Drummond
Drummond at avpvx1.ncifcrf.gov




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