[Histonet] endogenous peroxidases

[Rene J Buesa]

My sequence is: dewax (with dishwasher soap) → wash → HIER → wash → block endogenous peroxidase → go tthe IHC protocol.
René J.

--- On Fri, 4/22/11, sgoebel <@t> mirnarx.com <sgoebel <@t> mirnarx.com> wrote:


From: sgoebel <@t> mirnarx.com <sgoebel <@t> mirnarx.com>
Subject: RE: [Histonet] endogenous peroxidases
To: akemiat3377 <@t> yahoo.com, carl.hobbs <@t> kcl.ac.uk
Cc: histonet <@t> lists.utsouthwestern.edu
Date: Friday, April 22, 2011, 1:50 PM


So I have been seeing from this post that people block prior to HIER?  I
have always done it as the first step after for 10 minutes (I use the
DAKO enzyme blocker).  Is this wrong?
Thanks

Sarah Goebel, BA, HT(ASCP)
Histotechnologist
Mirna Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas  78744
(512)901-0900 ext. 6912


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Akemi
Allison
Sent: Friday, April 22, 2011 12:42 PM
To: Hobbs, Carl
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] endogenous peroxidases

Make sure you use a fairly fresh bottle of H2O2. (no older than 6  
months).  The stock bottle degrades after it is opened.  I have seen  
people using an older bottle of  H2O2 and the results are sub-optimal.
Akemi

Akemi Allison BS, HT (ASCP) HTL
Director
Phoenix Lab Consulting
Tele: 408.335.9994
E-Mail: akemiat3377 <@t> yahoo.com

On Apr 22, 2011, at 10:27 AM, Hobbs, Carl wrote:

>
> Gimme 5!
>
> Block before or after Ag retrieval.
> If it's Pwax sections make a 1/10 dilution of 100vols ( ~30%) H2O2  
> in Dist.water.
>  Block for 15mins.
> Methanol is not neccessary...imho.
> Carl
>
>
>
> Carl Hobbs
> Histology Manager
> Wolfson CARD
> School of Biomedical Sciences
> Kings College London
> Guys Campus
> London
> SE1 1UL
> Tel: 020 78486813
> Fax: 020 78486816
>
>
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