RE: [Histonet] Citrate Buffer pH

From:"Jeff Gordon"
Patsy, we have a steamer protocol for using our all in one deparaffinization and unmasking solutions Declere and Trilogy (US patent # 6,649,386) in a steamer.   We don't endorse it as often because we were looking to standardization and there were some antibodies that needed the pressure cooker heat to work optimally (such as ALK-1, Cyclin D1, CD10, and TTF-1) and so we chose to just continue to endorse using the pressure cooker.  The steamer one step method, however, works quite well, only it takes a bit longer (60 total minutes of steam time) than in the pressure cooker.  It is located on page 54 of the current Cell Marque reference guide.
 
Jeff Gordon
Cell Marque Corp.
-----Original Message-----
From: Patsy Ruegg [mailto:pruegg@colobio.com]
Sent: Wednesday, December 03, 2003 12:19 PM
To: Jackie.O'Connor@abbott.com; Pat Flannery
Cc: Histonet@lists.utsouthwestern.edu; histonet-admin@lists.utsouthwestern.edu
Subject: RE: [Histonet] Citrate Buffer pH

Has anybody tried the AR reagents that deparaffinzed such as BioCare Decloaker in a steamer in place of the pressure cooker?  PC is too harsh for some of my stuff but I like using these retrieval/deparaffination reagents.

Patsy

 

-----Original Message-----
From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Jackie.O'Connor@abbott.com
Sent: Tuesday, December 02, 2003 2:50 PM
To: Pat Flannery
Cc: Histonet@lists.utsouthwestern.edu; histonet-admin@lists.utsouthwestern.edu
Subject: Re: [Histonet] Citrate Buffer pH

 


I've been using a product from Biocare, a Universal Decloaker (6.0 citrate buffer) , which deparaffinizes during HIER in their pressure cooker (Decloaking Chamber) - they are a very Trekkie oriented company.  It works great - saves a bunch o' steps.  You might try contacting them for the specifics.  I don't think about the nature of these things if they work - I just follow along blindly . . . . . . . . .sometimes.

Jacqueline M. O'Connor HT(ASCP)
Abbott Laboratories
Global Pharmaceutical Research and Development
Discovery Chemotheraputics


 

Pat Flannery <pjfnefro@duke.edu>
Sent by: histonet-admin@lists.utsouthwestern.edu

12/02/2003 03:00 PM

       
        To:        Histonet@lists.utsouthwestern.edu
        cc:        
        Subject:        [Histonet] Citrate Buffer pH




I have a procedure for deparaffinizing (their word, not mine) tissue
sections in order to do immunostaining using the Vector MOM kit.  It
calls for heating the section (briefly) in a microwave in 0.1M Citrate
Buffer.  The procedure  does not specify the pH of the buffer, just to
make it by mixing 0.1M NaCitrate and 0.1M citric acid.  Does this sound
familiar to anyone out there?

--

-Patrick J. (Pat) Flannery
Division of Nephrology (that's kidneys to you)
Box 3014 (that's NOT "PO" just "Box")
Duke University Medical Center
Durham, NC  27710
E-mail: pjfnefro@duke.edu (preferred)
        FLANN002@MC.DUKE.EDU (also works)
Voice:   (919)660-6863
Fax:     (919)684-2929




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