[Histonet] Citrate Buffer pH

[Steven E. Slap]

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 <@t> bshsi.com
>
>
>-----Original Message-----
>From: Patsy Ruegg [mailto:pruegg <@t> colobio.com]
>Sent: Wednesday, December 03, 2003 1:19 PM
>To: Jackie.O'Connor <@t> abbott.com; Pat Flannery
>Cc: Histonet <@t> lists.utsouthwestern.edu; histonet-admin <@t> 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 <@t> lists.utsouthwestern.edu 
>[mailto:histonet-admin <@t> lists.utsouthwestern.edu]On 
>Behalf Of Jackie.O'Connor <@t> abbott.com
>Sent: Tuesday, December 02, 2003 2:50 PM
>To: Pat Flannery
>Cc: Histonet <@t> lists.utsouthwestern.edu; histonet-admin <@t> 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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