[Histonet] Methanol and CD markers

Gayle Callis gcallis <@t> montana.edu
Wed Jan 11 09:42:11 CST 2006

Just a note here. Methanol is known to create problems with CD marker 
immunohistochemistry and that is why many use hydrogen peroxide in buffer 
instead of this solvent.   This point was brought up in Jules Elias's book 
on immunohistochemistry and also on Histonet in the past. (see archives).

At 04:02 PM 1/10/2006, you wrote:
>We use both CD4 and CD8 on FFPE sections.  We had problems with the CD4 
>until we changed our quenching step to 0.5% H2O2 in MEOH for 10 min. The 
>pretreatment for both is 15 min microwave in pH 8 EDTA.
>Thomas Pier wrote:
>>I am looking for Reinhard von Wasielewski.  I recently saw a post
>>stating that he had working protocols for CD4 and CD8 that even worked
>>in decalcified bone marrow.  CD4 and CD8 in FFPE, decalcified bone
>>marrow is exactly what I'm having problems with.  I have been banging my
>>head against the wall with this for some time now.  I would really
>>appreciate any help I could get on this.  Thank you very much.
>>Tom Pier
>>Histonet mailing list
>>Histonet <@t> lists.utsouthwestern.edu
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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