[Histonet] restarting DAB reaction

[Tony Henwood]

Yes,

I would also agree that this would probably be the case.
I have not tried to redo the DAB step after counterstaining,
dehydrating, clearing and mounting.
I would expect it not to work.

Though I could easily be corrected.

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 




-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
jmyers1 <@t> aol.com
Sent: Wednesday, 15 October 2008 12:58 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] restarting DAB reaction


Emily:
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" <talulahgosh <@t> gmail.com>
Subject: [Histonet] restarting DAB reaction
To: histonet <@t> lists.utsouthwestern.edu

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?

Emily

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