ISH on fluorescing cells - (Apr/22/2012 )
Hey everyone, I want to perform in-situ hybridization on mRNA from cells that already express GFP and RFP. Will expression of these proteins interfere with the TSA fluorescence reaction that occurs during ISH? Is this why proteinase K is sometimes used so it destroys the fluorescing proteins?
Fixation before or during ISH usually destroys all fluorescence from GFP/RFP, so there should be no interference. PK is used to permeabilize your cells, but will certainly also destroy your GFP/RFP, so no problem at all.