Re: Need Antigen Retrieval Help
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From: | anita dudley <azdudley@hotmail.com> (by way of histonet) |
To: | histonet@histosearch.com |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
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
Dudley
Providence Hosp
Mobile, ala
>From: "Bennett, Catherine (Katie)" <cbennett@lrri.org>
>To: "'histonet@pathology.swmed.edu'" <histonet@pathology.swmed.edu>
>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
>in
>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
>appreciated.
>
>*********************************
>Catherine "Katie" Bresee Bennett
>Sr. Technical Associate
>Lovelace Respiratory Research Institute
>Albuquerque, New Mexico
>
>
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