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PFA Quenching in Staining of 2D and 3D MDCK Cultures - (Aug/31/2009 )

Hi! This is probably a stupid question...but what does is the purpose of "quenching" PFA after fixing cells to be analyzed via IF? What is the best quencher when using predominantly FITC and TxR secondary antibodies? How does this process of quenching work?

Thanks!!!

-carlymichelle-

PFA autofluoresces so it needs to be quenched so that you can tell actual signal from background. Sorry, I can't help you with the dye stuff, but the quenching is usually done using 0.1 M Glycine in several washes.

-bob1-