ELISA information..

crlong at vasci.umass.edu crlong at vasci.umass.edu
Mon Apr 3 12:28:28 EST 1995


	I would like to submit a question concerning antigen binding to ELISA
plates.  We are having difficulty obtaining any signal in our ELISA's when the 
same reagents give good response using dot blots to nitrocellulose.  Our
antigen is extracted using a 4M urea, 5% mercaptoethanol, 0.2% Triton X-100
buffer and the protein concentration is approx. 1.25 mg/ml.  Using this full
strength or diluting in different buffers with various pH and buffer
concentration has not been effective in generating a signal.  Different types
of plates have also been tried with no luck.  We are concerned that the
extraction buffer may be preventing the binding of the antigen to the plates,
is this likely?  We have attempted to dialyse the antigen against PBS and Tris
buffers and always get precipitation of the antigen.  Any information relavant 
to the problem would be greatly appreciated.

Thanks in advance,

Chuck Long



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