[Histonet] how to bleach melanoma cases without bad effect on IHC

[McMahon, Loralee A]

We have actually performed a bleach step before staining.  The only affect I have seen ( and we have only done a few antibodies) is that the tissue gets pretty chewed up.  Especially if there is a pretreatment step prior to the IHC staining.  

Loralee McMahon, HTL (ASCP)
Immunohistochemistry Supervisor
Strong Memorial Hospital
Department of Surgical Pathology
(585) 275-7210
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Cynthia Pyse [cpyse <@t> x-celllab.com]
Sent: Tuesday, March 22, 2011 10:01 AM
To: 'Shahram Sabeti'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] how to bleach melanoma cases without bad effect on IHC

Sabeti
I use the AEC chromogen (Dako catalog #K346989) for my melanoma cases. I use
a aqueous mounting media, placed on the tissue only, dry it completely on a
hot plate. When dry I coverslip it with permanent  mounting media(don't
place it in xylene), dry coverslip. Works great, and the color doesn't fade.

Cindy

Cindy Pyse, CLT, HT (ASCP)
Laboratory/Histology Supervisor
X-Cell Laboratories
716-250-9235
e-mail cpyse <@t> x-celllab.com



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Shahram
Sabeti
Sent: Tuesday, March 22, 2011 4:40 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] how to bleach melanoma cases without bad effect on IHC

hello dear fellows,
   i have an IHC project on paraffin embedded melanoma cases; some of them
being heavily pigmented. i have seen many protocols based on H2O2 to bleach
the pigmented ones, but i am not sure about any possible effect on antigen
retrieval.please guide me .
yours,
sabeti



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