Hi all
I was trying to perform a ELISA, in which I coat the plate with strep, then add in biotin conjugated peptide, then human serum and use anti-human IgG to detect antibody level to the peptide.
I found lots of my human serum samples have high IgG responses to streptavidin (determined by direct ELISA to strep). Is there a way to block these non-specific IgG binding to streptavidin to reduce my ELISA background?
Many thanks
Human sera are infamous for high background on immunosorbent assay platforms. Try adding 1M glucose, 0.1M glycine, and/or 1% non-human serum to your blocker.
One alternative is to use a small column containing Sephadex G10 bound to stepavidin or avidin and pass the human serum sample through it get rid off stepavidin interactive molecule. Use the eluted serum sample to perform ELISA.
I suppose you could try adding biotin to your blocker, to saturate the SA.
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