Need Antigen Retrieval Help

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From:"Bennett, Catherine (Katie)" <> (by way of histonet)
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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

Catherine "Katie" Bresee Bennett
Sr. Technical Associate
Lovelace Respiratory Research Institute
Albuquerque, New Mexico

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