Elisa - (Apr/22/2003 )

I'm doing a sandwich Elisa. I incubate the secondary Ab in the same solution that I dilute my samples with :1% BSA, 1% Casein, PBS .05%Tween-20. The assay Im doing is really sensitive and for the most doesnt seem to be working. Also, if Im using OPD how much stopping solution should I use?? Equal amount? Now I'm using 100uL OPD and 50uL stopping solution.
Thanks
Sharon

What do you mean of 'too sensitive'? Actually, ELISA is a rapid and sensitive method. I guess you mean the backgroud is over. Have you tried to use different combinations of Primary Ab and Secondary Ab at various concentration yet? To stop the reaction, I also used 50ul stopping solution.