[Histonet] RE: PIN4 antibody

Obregon, Cecilia CObregon <@t> mhs.net
Thu Aug 22 13:27:48 CDT 2013


We currently have the PIN4 optimized on both the Ventana XT and the Ultra using the IHC DS (Dual Stain) uDAB-uRed v3 procedure. 

       Std CC1 @37 degrees C, then K903/P63 Cocktail  incubation time 24 minutes, ultrawash
       Antibody denaturation 90 degrees C- 4minutes
       DS: 37degrees C, racemase (p504s) 36 minutes, ultrawash
       Finally, Hematoxylin and Bluing 4 minutes each. 

Antibody Information:
For the K903/P63 cocktail, we mix the 34BE12 (Ventana 790-4373) with the P63 (Biocare Cat#VP163G)
The P504S (P) is from Biocare Cat#VP200RG. 

Please beware of the new dehydration procedure if you're using the 'reformulated' alkaline phosphatase or Red Detection from Ventana as the chromogen is soluble in alcohol. Here's what we do once slides are finished: Rinse throughly in soapy DI water and bake for 10 minutes at 60 C (..you can also leave at room temperature for longer time I'd imagine), then 3 changes of xylene and coverslip as normal. 

Good luck!

       

________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Fawn Bomar [Fawn.Bomar <@t> HalifaxRegional.com]
Sent: Thursday, August 22, 2013 2:00 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] PIN4 antibody

Hi everyone,



Has anybody used the PIN4 cocktail using the Venatan Benchmark stainers?  If so, which company do you purchase it from and would you be willing to share your protocol?



Thanks



Fawn Bomar
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