THANKS!! request procedure for antigen retrieval with steam

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From:Connie McManus <>
To:Tom Wells <>,
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Histnett has come through once more...
I deeply appreciate all the great suggestions you have sent me on antigen
retrieval using steam and pressure cookers.  This is actually for my
husband who manages the Electron Microscope facilty on campus.  He was
doing an experiment using a type of IHC, but on tissues processed for EM.
They were having no luck seeing what they wanted to see (don't ask me what,
I'm NOT involved  *g*) and I thought maybe they should try antigen
retrieval.  I forget why using steam was better than doing a proteolytic
digest, but anyway,  that's the story.  

Thanks tons for your help!!

Connie McManus
Veterinary Diagnostics Lab
Utah State University
Logan, Utah

At 03:47 AM 06/24/2000 -0700, Tom Wells wrote:
>We use two methods, depending on the antigen:
>(1) Microwave pressure cooker. Partially fill the pressure cooker with water
>and place 2 plastic coplin jars filled with citrate buffer (but no slides)
>into the pressure cooker. We then preheat for approximately 15 minutes. Then
>remove the coplin jars and insert the slides. Continue heating for
>approximately 6 minutes. I then allow everything to cool for 15 minutes
>before proceeding with the method.
>(2)Autoclave method. For this method we use Dako target retrieval solution.
>Fill the autoclave with distilled water. Place the slides into plastic
>coplin jars and heat until the chamber reaches approximately 240 F. Continue
>heating for another 15 minutes. Once the chamber has returned to room
>temperature proceed with the method.
>Prior to removing the slides from the coplin jars I replace the heated
>buffer with room temperature distilled water. I hope this helps. Tom

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