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--></style><title>RE: [Histonet] Citrate Buffer
pH</title></head><body>
<div>Hi HistoNetters</div>
<div><br></div>
<div>As Dr. Shi points out in his book and articles about antigen
retrieval, there are three parameters involved in achieving
consistent results:</div>
<div><br></div>
<div>1) temperature</div>
<div>2) time</div>
<div>3) pH</div>
<div><br></div>
<div>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.</div>
<div><br></div>
<div>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.</div>
<div><br></div>
<div>best regards,</div>
<div>Steven Slap</div>
<div><br></div>
<div>At 3:53 PM -0500 12/3/03, Houston, Ronnie wrote:</div>
<blockquote type="cite" cite><font face="Arial" size="-1"
color="#0000FF">Patsy,</font></blockquote>
<blockquote type="cite" cite> </blockquote>
<blockquote type="cite" cite><font face="Arial" size="-1"
color="#0000FF">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.</font></blockquote>
<blockquote type="cite" cite> </blockquote>
<blockquote type="cite" cite><font face="Arial" size="-1"
color="#0000FF">Ronnie</font></blockquote>
<blockquote type="cite" cite> </blockquote>
<blockquote type="cite" cite><font face="Arial" size="-2"
color="#0000FF"><i>Ronnie Houston</i></font><i><br>
<font face="Arial" size="-2" color="#0000FF">Regional Histology
Operations Manager</font></i><i><br>
<font face="Arial" size="-2" color="#0000FF">Bon Secours
HealthPartners Laboratories</font></i><i><br>
<font face="Arial" size="-2" color="#0000FF">5801 Bremo
Road</font></i><i><br>
<font face="Arial" size="-2" color="#0000FF">Richmond, VA
23226</font></i><i><br>
<font face="Arial" size="-2" color="#0000FF">(804) 287
7972</font></i><i><br>
<font face="Arial" size="-2"
color="#0000FF">ronnie_houston@bshsi.com</font></i></blockquote>
<blockquote type="cite" cite> <br>
<blockquote><font size="-1">-----Original Message-----<br>
<b>From:</b> Patsy Ruegg [mailto:pruegg@colobio.com]<br>
<b>Sent:</b> Wednesday, December 03, 2003 1:19 PM<br>
<b>To:</b> Jackie.O'Connor@abbott.com; Pat Flannery<br>
<b>Cc:</b> Histonet@lists.utsouthwestern.edu;
histonet-admin@lists.utsouthwestern.edu<br>
<b>Subject:</b> RE: [Histonet] Citrate Buffer pH</font><br>
<font size="-1"></font></blockquote>
<blockquote><font face="Arial" size="-1" color="#000080">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.</font></blockquote>
<blockquote><font face="Arial" size="-1"
color="#000080">Patsy</font></blockquote>
<blockquote><font face="Arial" size="-1"
color="#000080"> </font></blockquote>
<blockquote><font size="-1" color="#000000">-----Original
Message-----<br>
<b>From:</b> histonet-admin@lists.utsouthwestern.edu
[mailto:histonet-admin@lists.utsouthwestern.edu]<b>On Behalf Of</b>
Jackie.O'Connor@abbott.com<br>
<b>Sent:</b> Tuesday, December 02, 2003 2:50 PM<br>
<b>To:</b> Pat Flannery<br>
<b>Cc:</b> Histonet@lists.utsouthwestern.edu;
histonet-admin@lists.utsouthwestern.edu<br>
<b>Subject:</b> Re: [Histonet] Citrate Buffer pH</font></blockquote>
<blockquote><font face="Times New Roman"> </font></blockquote>
<blockquote><font face="Times New Roman" color="#000000"><br>
</font><font face="Arial" size="-1" color="#000000">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.</font><font color="#000000"><br>
<br>
<font face="Arial" size="-1">Jacqueline M. O'Connor HT(ASCP)<br>
Abbott Laboratories<br>
Global Pharmaceutical Research and
Development</font></font></blockquote>
<blockquote><font face="Arial" size="-1" color="#000000">Discovery
Chemotheraputics</font></blockquote>
</blockquote>
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