Re: Need Antigen Retrieval Help

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From:anita dudley <> (by way of histonet)
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Katie,  when I do my antigen retrieval I use a 25 slide holder full of
slides, blank slides where needed.  I make 200cc of retrival solution, fill
a plastic staining container and microwave the slides for 5 mins on high, I
then add 50cc of water to the solution and microwave 5 more mins.  Then they
sit on the counter for 25 mins. rinse for 5 mins and they are ready for the
antiboby.  I use Biogenex antigen retrieval.  Hope this helps.   Anita
                                       Providence Hosp
                                       Mobile, ala

>From: "Bennett, Catherine (Katie)" <>
>To: "''" <>
>Subject: Need Antigen Retrieval Help
>Date: Thu, 11 Nov 1999 09:20:20 -0700
>Hi all!
>I'm starting a double staining IHC run this morning that is intending to
>take two days, and I may have already committed a fatal error on step one,
>so I'm wondering if I should continue or start over.
>I am doing microwave antigen retrieval and had a problem with the solution
>boiling over and completely out of the coplan jar.  My protocol calls for
>microvaving slides in a coplan jar of antigen retrieval for 2x 5 min, but
>after just 1.5 min all the solution is boiled over and gone.  Needless to
>say, I didn't catch it in time and the slides may have gone through some
>microwaving without any solution in the jar.  I had placed the coplan jar
>a small dish of shallow water, so the microwave was very humid at least.
>I'm guessing the microwave is much more powerful than the one used in the
>protocol I am using and I should experiment with which power setting I
>should use.  However, in the meantime, are the slides from today toast
>because they have been "cooked" without being in the antigen retrieval
>solution, or can I continue with them?
>Any suggestions from others on preventing boil-over would also be much
>Catherine "Katie" Bresee Bennett
>Sr. Technical Associate
>Lovelace Respiratory Research Institute
>Albuquerque, New Mexico

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