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From:Katri Tuomala <> (by way of histonet)
To:histonet <>
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Mary L. Verlinde wrote:
> I am looking for a recipe for an antigen retrieval acid or buffer for
> paraffin slides to do IHC. Thanks in advance.
> Mary Verlinde
> Animal Health Diagnostic Lab
Hi Mary,
Citrate buffer pH 6.0 is commonly used in microwave retrieval of
formalin fixed, paraffin embedded material. I make my own by dissolving
2.1 g of citric acid in 1000 ml dist.water and bringing the pH up to 6.0
by adding 2M sodium hydroxide. I use microwavable pressure cooker, which
allows continued heating without excessive evaporation of the buffer.
Bring the slides in buffer to boiling 10-15 minutes on HIGH. Immediately
add another 2-3 minutes on HIGH. Then let slides sit in hot solution for
20 minutes. Continue with your method. You can do this in coplin jars,
if you don't have a pressure cooker, but you have to add more buffer
every 5 minute intervals of heating to avoid the slides from drying out.
To obtain reproducible results try to keep the amount of the solution
and number of slides constant. Hope this is helpful, good luck! Katri.

Katri Tuomala
Department of Pathology
St.Joseph's Hospital
Hamilton, Ontario, Canada

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