[Histonet] RE: Trypsin versus other AR-s

[Swain, Frances L]

I use Trypsin only as a last resort.  I use Pepsin because it is more gentle (does not chew up the sections).  I do not use Proteinase K unless I absolutely have to and for just a few minutes as it will eat the sections off of the slides.  If I use Citrate Buffer I heat it up in the microwave first then quickly put the slides in it cover it and let it sit on the bench top until cool about 20 minutes.  As to why we use Citrate buffer there has been several discussions about that on the histonet.
You might want to search the archieves and see what others have had to say.

Frances L. Swain HT(ASCP) A. A. S.
Special Procedures Technician
Department of Orthopaedic Surgery
Center for Orthopaedic Research
Barton Research Building 2R28
4301 West Markham Street
Little Rock AR 72205
(501) 686-8739 PHONE
(501) 686-8987 FAX
swainfrancesl <@t> uams.edu email

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Margaryan, Naira
Sent: Friday, March 27, 2009 2:38 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Trypsin versus other AR-s

Hi Dears,

 

It is Friday, but not a weekend yet:-):-(

 

I just got a request from my PI to figure out:

1.      How often now day's people use Trypsin (EDTA, Proteinase K or E)
as an Antigen Retrieval for FFPE. 

2.      Why or is the Citrate Buffer pH6 more usable??? 

3.      Is Trypsin very old technique? 

 

Any educational feedback is appreciated,

Naira

 

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