Re: Need Antigen Retrieval Help

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From:Mary Latimer <> (by way of histonet)
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Dear Katie  I have no personel experience of this technique but on an
immuno course That I attended in London  at the Hammersmith Hospital  when
these techniques were being discussed one of the techs there said that she
did these in an autoclave rather than microwave her reasons were that the
microwave needed constant watching and the addition of near boiling
solution to avoid drying out this apparently is not the case if you use an
autoclave or pressure cooker there is a written method in a book called
Introduction to Immunocytochemistry by Polak and Van Noorden I can fax it
if you would like to see it please email ....Hope this Helps Mary

On Thu, 11 Nov 1999, Bennett, Catherine (Katie) wrote:

> 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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