[Immunology] Elispot

Catalin Tucureanu via immuno%40net.bio.net (by catalintucureanu from gmail.com)
Mon Apr 14 05:51:40 EST 2008

 We are having some problems setting up an murine IFN-gamma Elispot  
assay (using an antibody pair and streptavidin-HRP from R&D that worked 
verry well for ELISA and MAIPS4510 plates from Millipore). Could anybody 
explain why we are getting spots (proportional in number with the IFN 
concentration) even in the wells where we have used the recombinant 
protein as a positive control for the antigen antibody reaction 
(shouldn't the membranes be uniformly coloured instead, given the fact 
the antibody should have coated the entire membrane and the protein is 
in a homogenous solution free to bind any antibody it comes across).  
Any information is wellcome!

Thank you !
Catalin Tucureanu

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