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cellular ELISA - cellular ELISA involving determing Ab concentration (May/16/2007 )

Hi, I need to determine the concentration of Abs which ware purified from hybridoma.

how do I develope an ELISA protocol for measuring the concentration of a mAb for a cell surface marker (such as CD4 or CD8, and I didnt purified the proteins)?
with what do I coat the plate in that case?

Does anybody have a suggestion?


well, what you are thinking of is perhaps more of a functionality test

if you are only wanting to know your antibody concentration, in what was your hybridoma raised? for example, if your hybridoma was raised in mouse, then you could use an anti-mouse antibody in your elisa

another option is to take your purified antibody and spec it, like any other protein