antigen retrieval using steamer

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From:"Kathy Oprea" <koprea@calc.vet.uga.edu>
To:HistoNet@Pathology.swmed.edu
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Date:Tue, 18 May 1999 11:52:51 EST
Content-Type:text/plain; charset=US-ASCII

Our lab has been routinely using the steamer for antigen retrieval 
for 3 years with consistently good results. Our procedure is very 
simple- We cut 4 micron sections for IHC. Fill steamer, with tap 
water, and start. Deparaffinize slides in xylene, hydrate through 
graded ETOH, to deionized water. Drain water from slides and place 
them in citrate buffer. (We currently use BioGenex Antigen Retrieval 
Citra, Catalog# HK086-9K. This comes in a 10X concentrate which we 
dilute for a working solution,  following manufacturers directions.) 
Place slides/citrate buffer in steam chamber base. Cover 
and steam for 20 minutes. Carefully remove covered steam chamber base 
containing slides/citrate buffer, from steamer. Open the lid and let 
slides/citrate buffer, remain inside steam chamber base for an 
additional 5 mintes to cool. Proceed with routine IHC.   
Feel free to contact me with questions.
Kathy 
************ Thank you ************
* Kathy Oprea                     *
* University of Georgia           *
* College of Veterinary Medicine  *
* Athens, GA 30602-7388           *
* KOPREA@calc.vet.uga.edu         *
The worker who's never said "Thank
God it's Friday," probably never
really worked.
************************************



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