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
