RE: HIER Methods Survey

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From:"Nocito, Joseph" <>
To:'Luis Chiriboga' <>, Histonet <>

1. most of our antibodies are steamed, some need to be enzymatically
digested with Proteinase K, Pepsin or Trypsin.
2.  We steam for 20 minutes and cool for 20 minutes
3. We use the Dako Autostainer
4.  We use Dako's Antigen Retrieval Solution, and Biocare's EDTA, Nuclear
Decloaker, Antigen Decloaker and Reveal.
Different antibodies require different methods.  I read the specificity
sheet to determine the method to use.  But, the Hier methods are dependent
on your fixation and processing.  what works for me might not work for you.

Joe Nocito, B.S., HT(ASCP)QIHC
Histology Supervisor
Christus Santa Rosa Hospitals
San Antonio, Texas 

> -----Original Message-----
> From:	Luis Chiriboga []
> Sent:	Wednesday, May 31, 2000 8:35 AM
> To:	Histonet
> Subject:	HIER Methods Survey
> Dear Histonetters:
> I am taking an informal survey to see what HIER methods/techniques
> everyone is using.  As with any survey,  the more responses the better.
> If anyone has additional questions they would like to add please e-mail
> me directly and i will reissue a questionnaire.
> 1) Which heating method do you use for HIER? (i.e.. Microwave, pressure
> cooker, combination etc...)
> 2) What is the average "real" heating time for antigen retrieval?
> (actual boiling time)
> 4) How long is your cooling period?
> 5) Do you perform manual or automated IHC?
> 5) Do you prepare your own retrieval solutions or purchase commercial?
> 6) Do you use more than one HIER solution? (please list)
> 7) Which solution do you use the most?
> I will post the stats as soon as I compile all the responses.
> Thanks in advance
> --
> Luis Chiriboga Ph.D HT(ASCP)QIHC
> Department of Pathology 4W27
> Bellevue Hospital
> NYU School of Medicine
> First Avenue & 27th Street
> New York, NY 10016
> (212)562-4667

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