RE: [Histonet] restarting DAB reaction

Although it might seem that the reapplication of a substrate-chromogen solution?after removal of the coverslip/mounting media and rehydration?should work, it?very rarely does.? The scientific reason is that, after the specimen has been dehydrated and coverslipped,?the enzyme label (e.g. polymer-HRP) applied in the original staining procedure is 'dead' -- meaning that it lacks the?power to drive another chromogen-polymerization reaction.? Your best bet is start over, 'from scratch'...
Good Luck,

J.D.?Myers, M.S., CT(ASCP)


Message: 1
Date: Mon, 13 Oct 2008 13:17:49 -0400
From: "Emily Sours" 
Subject: [Histonet] restarting DAB reaction

Has anyone ever tried to continue a DAB reaction after the slides have been
washed and coverslipped?
I can't think of a reason why this wouldn't work, since we just wash the
slides in PBS and water after the DAB step.
We use a vectastain kit for this--would we need to add more of the ABC
reagent (after removing the coverslips and washing in PBS) and then do DAB?


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