Re: request procedure for antigen retrieval with steam

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From:Dana Settembre <settembr@UMDNJ.EDU>
To:Connie McManus <conmac@cc.usu.edu>
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On Fri, 23 Jun 2000, Connie McManus wrote:

>  Hi gang!
> 
> what are the procedures you use with your steamers and pressure cookers?
> Although I need this for plastic embedded 1 um sections for electron
> microscopy, whatever procedures you use, I am interested in.  
> 
> Thanks...
> 
> Connie McManus
> Veterinary Diagnostics Lab
> Utah State University
> Logan, Utah
> USA
> 
Connie
After I deparaffinize running the slides down to water I use Target
Retrieval Solution from Dako Cat# S1699.  This is a 10X concentration that
you are to use at a 1:10 dilution.  They also have it in a Ready to Use 
solution.  I set up the steamer at least 30 minutes before hand, put the
solution in a plastic coplin jar and place the coplin jar in the
center of the steamer, if I have one coplin jar, or evenly spaced out in
the steamer if there are more than one coplin jars.  I set the steamers
timer for 30 minute pre-heating.  After pre-heating I place the slides in
the hot solution and incubate for 30 minutes.  Then I (carefully) pull the
coplin jar(s) out of the steamer and cool down for 20 minutes.  Then place
slides into buffer and continue with protocol.


Dana Settembre
Immunohistochemistry Lab
Pathology Department
University Hospital
Newark,  New Jersey
USA




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