Patsy,

In my experience, staining quality decreases as the time between HIER and
staining increases.  I believe the epitopes you retrieve with heat do not
stay open indefinitely and will, in fact, begin to slowly close very shortly
after HIER.  So, for the best staining, I would recommend no wait time at
all between retrieval and staining.

Do you have a tech that comes in earlier than you?  Perhaps they could begin
the 3 hr. water bath incubation for you before you get in so you don't have
to wait around.  I have done this myself and it works well if the early tech
is available.

Good Luck,

Glen Dawson BS, HT & QIHC (ASCP)
IHC Manager
Milwaukee, WI

-----Original Message-----
From: Patsy Ruegg [mailto:pruegg@ihctech.net]
Sent: Thursday, February 17, 2005 2:33 PM
To: ihcrg@neo.agsci.colostate.edu
Cc: histonet@pathology.swmed.edu
Subject: [Histonet] antigen retrieval question


I have heard that it is not a good idea to do HIER one day and do the IHC
the next?  Is this so?  I heard that the unmasking could actually remask if
left sitting a long time before doing the staining.  I have a protocol that
requires a 3 hr. water bath incubation for AR and it would be more
convienent if I could do that one day and do the rest of the procedure the
next.
Patsy
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