problem with an ELISA
rczci at unix.ccc.nottingham.ac.uk
Thu Jan 6 12:19:39 EST 2000
> Christiane Devaux wrote:
> > What did I have to change to clean my results ? I read a paper about
> > dilution of plasma but I don't think it's a good idear. Is there another
> > solution ???
> A typical problem is the presence of rheumatoid factor in sera:
> antibodies that bind the Fc of other antibodies. This crosslinks your
> detecting antibody with the capture antibody, giving "Background". The
> best way to avoid this is to use a F(ab)2 fragment for either capture
> or detection. Yes, it's more expensive (and harder to find
> commercially), but well worth it in terms of data quality.
> Good luck,
You can also just include non-immune anti-sera (bog standard antibody that
isn't going to react with anything) into the buffer with the detecting
antibody before/during application. Much cheaper.
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