RE: EM IHC questions

From:Cynthia Favara

I am not knowledgeable in this area but know there are many publications on
the subject. I would go to PUBMED and run a lit search. This should give you
what is currently being done with protocols. If protocols are not listed get
in touch with the author. They are usually very helpful.

Cynthia Favara
903 South 4th Street
Hamilton, MT 59840

-----Original Message-----
From: marjorie lehman []
Sent: Wednesday, October 30, 2002 7:34 AM
To: histonet
Subject: EM IHC questions

Dear EM Netters,
Here I am again with questions. I have been asked to help the EM people with
immuno electron histochemistry, and this is a "Halt leading the Blind"
situation! I did EM work back in the middle ages, but did not do any EM IHC
(there was no such thing), so I am somewhat familiar with EM. The EM tech is
not familiar with any IHC whatsoever.
They are working with human skin.
I have finally convinced them not to fix in gluteraldehyde, although they
insist on going with 1% gluter in paraformaldehyde, and to use low temp
(Lowcryl) processing. I gave them a K15 Ab which I use with great success
which I figured would stain lots of basal cells at least. I also gave them
method and suggested they bracket their dilutions and incubate overnight.
They got no reaction.
I am aware that not all Abs work with EM sections, obviously I don't know
ones do/don't! So the 1st question is: can anyone tell me which don't and/or
least tell me one that does. Since I don't know whether their method is the
problem or the K15 just won't work, I can't trouble shoot.
The 2nd question is: can one do HIER (pressure cooker) and or enzyme
on EM sections?
Thanks for any help/advise anyone can offer.
Marge Lehman 

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