Elispot Discussion

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Elispot Discussion

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Elispot Discussion: Please, feel free to start discussions about the Elispot technique, methods, ideas, questions, experience etc. With your input, we would like to make this website to a special interactive place for everyone who is interested in the Elispot assay.

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Subject Blocking

Date Wed Nov 14 2007 18:35

Author Retro (bjorghi@hi.is)



Hi there! I'm a beginner in this ELISPOT thing and just gaining some experience by doing some simple tricks. Recently I've been using B-cells, fresh from buffy coat and EBV transformed B-cells. I find it difficult to trust this method because in my control where I did a blocking step and a secondary antibody step (no primary antibody step), I got a lot of background and small spots, similar to those seen in wells with cells in it. This must indicate that the blocking step has failed (?). What kind of blocking material do you use (I've used 5% BSA in PBS) and has anyone tried to incubate with AIM-V instead of RPMI + serum ??

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