Trisha,
I have not done this specifically; however, it should be possible to combine
the two techniques. The effectiveness of double staining or re-staining the
BrdU slides may depend on the method used to denature the DNA before
staining with Anti-BrdU (acid, endonuclease, etc).  The apoptotic cells
probably did not incorporate much BrdU so TUNEL or perhaps easier the
Anti-PARP p85 Fragment pAb from Promega could be used as a marker for
apoptosis.

Ty Lee


-----Original Message-----
From: P. Emry <emry@u.washington.edu>
To: Technical Services <techserv@dakousa.com>
Cc: Cindy Chard-Bergstrom <CHARD_B@vet.ksu.edu>;
Histonet@Pathology.swmed.edu <Histonet@Pathology.swmed.edu>
Date: Tuesday, April 27, 1999 3:33 PM
Subject: Re: Double Staining


>Hi,
>I have an over eager visiting fireman...he did Brdu on slides meant for
>apopotosis.  I am sure this is the dumbest question yet...but is it
>possible to re-stain or combine the two...ok...go ahead and laugh.  Wish I
>knew how to make this anonymous.
>Trisha
>On Tue, 27 Apr 1999, Technical Services wrote:
>
>> Hi Cindy.
>>
>> We have a double-stain kit at DAKO which utilizes our Envision polymer
>> system, and includes a blocking reagent that enables you to completely
carry
>> out one stain, eliminate any vestige of it (aside from the colorimetric
>> reaction of course) and then proceed with another stain. You can use two
>> polyclonals, two monoclonals, what have you. I've spoken with people who
>> have even quadruple stained. If you would like more information please
>> contact me.
>>
>> Thank you,
>>
>> Joel Weisenberger
>> DAKO Corporation
>> Technical Services
>> 800-235-5743 x5325
>> techserv@dakousa.com
>>
>>
>> -----Original Message-----
>> From: Cindy Chard-Bergstrom <CHARD_B@vet.ksu.edu>
>> To: Histonet@Pathology.swmed.edu <Histonet@Pathology.swmed.edu>
>> Date: Tuesday, April 27, 1999 9:59 AM
>> Subject: Double Staining
>>
>>
>> >Dear Histonetters,
>> >
>> >    I will soon be working on a double staining IHC project. I seem
>> >to remember someone discussing a proceedure that involved stripping
>> >off the first primary, after addition of the chromogen, and then
>> >applying the second primary. Does anyone have this protocol or any
>> >others involving double staining. One project involves two rabbit
>> >primary antibodies and there is not enough antibody to conjugate
>> >either one to HRPO or alk-phos. Thanks to all in advance.
>> >
>> >Cindy Chard-Bergstrom, BS, HT(ASCP)
>> >
>> >
>> >
>> >
>> >
>>
>>
>>
>
>Trisha Emry
>U of Washington, Seattle
>
>
>