[Histonet] Re: Which one first - antigen retrieval or peroxidaseblock

[Patsy Ruegg]

Again this is all antigen dependent.  CD4 is one that I have found does
better if you do endogenous enzyme block after AR and for that one I also
make sure I use h202 in methanol, so it may be the fixing action of the
methanol as much as the blocking of the h202 at play here.

Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. #215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
pruegg <@t> ihctech.net
www.ihctech.net
www.ihcrg.org    
 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Tuesday, September 30, 2008 6:51 AM
To: AGrobe2555 <@t> aol.com; histonet <@t> lists.utsouthwestern.edu; Greg Dobbin
Subject: Re: [Histonet] Re: Which one first - antigen retrieval or
peroxidaseblock

As far as I remember enzymes are proteins and are they not supposed to
cross-link with formalin also?
René J.

--- On Mon, 9/29/08, Greg Dobbin <gvdobbin <@t> ihis.org> wrote:

From: Greg Dobbin <gvdobbin <@t> ihis.org>
Subject: Re: [Histonet] Re: Which one first - antigen retrieval or
peroxidase block
To: AGrobe2555 <@t> aol.com, histonet <@t> lists.utsouthwestern.edu
Date: Monday, September 29, 2008, 10:12 PM

Peroxidase is an enzyme. It doesn't miraculously appear as a result to
exposure to heat.  You show me one one slide where this has occurred and
I will humbly remain silent in this forum for a whole month! Ok, maybe
just a week...  :-)
Greg

Greg Dobbin, R.T.
Chief Technologist, Anatomic Pathology
Dept. of Laboratory Medicine,
Queen Elizabeth Hospital,
P.O. Box 6600
Charlottetown, PE    C1A 8T5
Phone: (902) 894-2337
Fax: (902) 894-2385

"And in the end it's not the years in your life
that count. It's the life in your years." 
- Abraham Lincoln

>>> <AGrobe2555 <@t> aol.com> 09/29/08 5:59 PM >>>
 
I'm with Rene on this one.  Block after retrieval.
 
For those of you who do the block before retrieval, how do you  control
for 
any activity that might be exposed during the retrieval  step?  Couldn't
it 
lead to false positives in your slides? 
 
Albert C.  Grobe, PhD
Tissue Engineering Lab
International Heart Institute of Montana  Foundation




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