Fc block/non-specific block

From: Kennedy, E. (Elaine) (E.Kennedy@organon.nhe.akzonobel.nl)
Date: Tue Apr 16 2002 - 09:13:39 EST


Hi all,

I'm trying to detect an intracellular antigen on a monocytic cell line using
an unlabelled primary followed by a RPE labelled seconday so am anticipating
a lot of background.

I know I have to block Fc receptors to reduce background but can I do this
after my fixation/permeabilisation step?

Also, I come from a immunohistochemistry background it makes sense to me to
also use a blocking step which caters for non-specific binding of the
antibody - is it possible to do this (perhaps in conjunction with the Fc
blocking step)?

I'd appreciate any tips.

Thanks,
Elaine Kennedy
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