[Histonet] H2O2 on CD markers

[Gayle Callis]

>Date: Mon, 29 Mar 2004 08:40:21 -0700
>To: Amos Brooks <amosbrooks <@t> earthlink.net>
>From: Gayle Callis <gcallis <@t> montana.edu>
>Subject: Re: [Histonet] H2O2 on CD markers
>In-Reply-To: <6.0.0.22.0.20040327190223.01b9ede8 <@t> pop.earthlink.net>
>References: <E1B7I6G-00031X-Pp <@t> swlx162.swmed.edu>
<E1B7I6G-00031X-Pp <@t> swlx162.swmed.edu>
>
>Amos,
>
>Are you working with human or mouse tissues?  Murine CD4 and CD8 is a
total failure with formalin fixed paraffin tissues, and no retrieval or
digestion recovers these antigens. They do work, however with zinc TRIS
buffer fixation where no retrieval is needed, but endogenous peroxidase
cannot be successfully/totally blocked with this fixation.  Alkaline
phosphatase methods work well with ZnTRIS to avoid peroxidase blocking.
This is why we stay with frozen sections for murine CD4 and CD8.
>
>
>At 07:07 PM 3/27/2004 -0600, you wrote:
>>Jorge,
>>         I don't have any rule of thumb to go by, merely evidence that 
>>there is an effect. Our CD4 works best if you do the H2O2 block prior to Hi 
>>pH antigen retrieval (pH 10.0 vs 7.6). In fact if you forget and block 
>>after retrieval it may not label at all. It's worth doing some 
>>experimentation on.
>>Amos Brooks
>>
>>At 12:00 PM 3/27/2004, you wrote:
>>>Hello everybody.
>>>I have a simple question: do someone know the effect, if any, of the
>>>different concentrations, and times, of H2O2 on the CD markers (Paraffin
>>>and/or frozen sections)? I have read that some people use 0.01%, others
0.1%
>>>and others 3% for different times, usually 10 or 30 minutes. What could
>>>happen if I use a longer time?
>>>Thank You very much.
>>>
>>>JORGE IVAN ZAPATA
>>
>>
>>
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>>