RE: [Histonet] Citrate Buffer pH
RE: [Histonet] Citrate Buffer
pH
Hi HistoNetters
As Dr. Shi points out in his book and articles about antigen
retrieval, there are three parameters involved in achieving
consistent results:
1) temperature
2) time
3) pH
How you get there (steamer, water bath, microwave, pressure
cooker) doesn't matter, so long as you can successfully monitor these
three variables very accurately.
As Jeff Gordon correctly points out, some antigens need
temperatures higher than 98°C for optimal retrieval, however.
For this reason, pressure cookers, whether by themselves, or inside of
a microwave have been used to reach temperatures as high as 120°C as
needed. Using a combination microwave/pressure method, I have
been able to use citrate buffer at pH 6.0 to retrieve most antigens at
110°C in 3-5 minutes.
best regards,
Steven Slap
At 3:53 PM -0500 12/3/03, Houston, Ronnie wrote:
Patsy,
We routinely use Reveal and Borg, both from Biocare
Medical, with great success in a B&D steamer, 20 minutes,
followed by 2 rinses in hot buffer, and 20 minute cool down. Very
rarely use pressure cooker, and don't use micrwave at
all.
Ronnie
Ronnie Houston
Regional Histology
Operations Manager
Bon Secours
HealthPartners Laboratories
5801 Bremo
Road
Richmond, VA
23226
(804) 287
7972
ronnie_houston@bshsi.com
-----Original Message-----
From: Patsy Ruegg [mailto:pruegg@colobio.com]
Sent: Wednesday, December 03, 2003 1:19 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
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