Re: antigen retrieval

From:"Johnson, Theresa"

Veronique,
In the past I have used the microwave (over-the-counter model) almost
exclusively for antigen retrieval.  I have used a rice steamer as well, and
while it's slower, it gets good results.  Currently I have a pressure cooker
model from BioCare Medical and am very happy with the results.  The
investigators at this facility looked mortified when I mentioned microwave
antigen retrieval, worried that their proteins would be adversely affected
by the action of microwaves.  The electric pressure cooker was a solution
they could live with.  It also seems to answer some of the questions of
quality control with regard to including steam monitor strips in each run.
As for a book recommendation, an excellent text on the subject is Antigen
Retrieval Techniques: Immunohistochemistry and Molecular Morphology, ed. by
Shi, Gu, and Taylor, ISBN 1-881299-43-0.  I got my particular copy at
Amazon.com.
Good luck to you!
Teri Johnson
Manager Histology Core Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


hi everybody ! Being pretty new in the big world of immunohistochemistry I
am asking myself lots of questions (some people tells me it's a good
thing..). I know that I should get a good book on the subject (any advice ?)
but I find that "real-life" experience is often much more informative. So
the question of the day is about antigen retrieval. I have been looking
around for different protocols and most of them use a microwave oven. I
can't buy right now a microwave especially for IHC and I am a bit concerned
about setting up parameters on an oven used extensively for other purposes.
I am a bit anxious that the life span of this oven is going to be pretty
short... So, I tried the heating plate protocol (10mn in boiling citrate
buffer) on brain sections. So far, it seems to be working pretty well. But I
may be lucky. I am therefore wondering if people out there tried this and
what you think about it. What make you prefer the microwave method and here
is the ultimate question : is a microwave oven a "must have" for doing
incredible IHC ? all your advices are welcome... veronique. 







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