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<DIV><SPAN class=130271719-03122003><FONT face=Arial color=#0000ff size=2>Patsy,
we have a steamer protocol for using our all in one<SPAN
class=320542719-03122003> deparaffinization and unmasking</SPAN> solutions
Declere and Trilogy<SPAN class=320542719-03122003> (US patent
# </SPAN><FONT face="Times New Roman"><SPAN class=320542719-03122003><SPAN
class=320542719-03122003><FONT face=Arial>6,649,386)
</FONT></SPAN></SPAN></FONT>in a steamer. <SPAN
class=320542719-03122003> </SPAN>We don't endorse it a<SPAN
class=320542719-03122003>s often</SPAN> 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, <SPAN
class=320542719-03122003>CD10, </SPAN>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 <SPAN class=320542719-03122003>reference
guide</SPAN>.</FONT></SPAN></DIV>
<DIV><SPAN class=130271719-03122003><FONT face=Arial color=#0000ff
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=130271719-03122003><FONT face=Arial color=#0000ff size=2>Jeff
Gordon</FONT></SPAN></DIV>
<DIV><SPAN class=130271719-03122003><FONT face=Arial color=#0000ff size=2>Cell
Marque Corp.</FONT></SPAN></DIV></DIV>
<BLOCKQUOTE dir=ltr style="MARGIN-RIGHT: 0px">
<DIV class=OutlookMessageHeader dir=ltr align=left><FONT face=Tahoma
size=2>-----Original Message-----<BR><B>From:</B> Patsy Ruegg
[mailto:pruegg@colobio.com]<BR><B>Sent:</B> Wednesday, December 03, 2003 12:19
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<BR><BR></FONT></DIV>
<DIV class=Section1>
<P class=MsoNormal><SPAN class=EmailStyle16><FONT face=Arial color=navy
size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Arial; mso-bidi-font-size: 12.0pt">Has
anybody tried the AR reagents that deparaffinzed such as BioCare Decloaker in
a steamer in place of the pressure cooker?<SPAN
style="mso-spacerun: yes"> </SPAN>PC is too harsh for some of my stuff
but I like using these retrieval/deparaffination
reagents.<o:p></o:p></SPAN></FONT></SPAN></P>
<P class=MsoNormal><SPAN class=EmailStyle16><FONT face=Arial color=navy
size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Arial; mso-bidi-font-size: 12.0pt">Patsy<o:p></o:p></SPAN></FONT></SPAN></P>
<P class=MsoNormal><SPAN class=EmailStyle16><FONT face=Arial color=navy
size=2><SPAN
style="FONT-SIZE: 10pt; FONT-FAMILY: Arial; mso-bidi-font-size: 12.0pt"><![if !supportEmptyParas]><![endif]> <o:p></o:p></SPAN></FONT></SPAN></P>
<P class=MsoNormal style="MARGIN-LEFT: 0.5in"><FONT face=Tahoma color=black
size=2><SPAN
style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Tahoma">-----Original
Message-----<BR><B><SPAN style="FONT-WEIGHT: bold">From:</SPAN></B>
histonet-admin@lists.utsouthwestern.edu
[mailto:histonet-admin@lists.utsouthwestern.edu]<B><SPAN
style="FONT-WEIGHT: bold">On Behalf Of
</SPAN></B>Jackie.O'Connor@abbott.com<BR><B><SPAN
style="FONT-WEIGHT: bold">Sent:</SPAN></B> Tuesday, December 02, 2003 2:50
PM<BR><B><SPAN style="FONT-WEIGHT: bold">To:</SPAN></B> Pat
Flannery<BR><B><SPAN style="FONT-WEIGHT: bold">Cc:</SPAN></B>
Histonet@lists.utsouthwestern.edu;
histonet-admin@lists.utsouthwestern.edu<BR><B><SPAN
style="FONT-WEIGHT: bold">Subject:</SPAN></B> Re: [Histonet] Citrate Buffer
pH</SPAN></FONT></P>
<P class=MsoNormal style="MARGIN-LEFT: 0.5in"><FONT face="Times New Roman"
size=3><SPAN
style="FONT-SIZE: 12pt"><![if !supportEmptyParas]><![endif]> <o:p></o:p></SPAN></FONT></P>
<P class=MsoNormal
style="MARGIN-BOTTOM: 12pt; MARGIN-LEFT: 0.5in; MARGIN-RIGHT: 0in; mso-margin-top-alt: 0in"><FONT
face="Times New Roman" color=black size=3><SPAN
style="FONT-SIZE: 12pt; COLOR: black"><BR></SPAN></FONT><FONT face=Arial
color=black size=2><SPAN
style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Arial">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.</SPAN></FONT><FONT color=black><SPAN
style="COLOR: black"> <BR><BR></SPAN></FONT><FONT face=Arial color=black
size=2><SPAN
style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Arial">Jacqueline M.
O'Connor HT(ASCP)<BR>Abbott Laboratories<BR>Global Pharmaceutical Research and
Development<BR>Discovery Chemotheraputics<BR></SPAN></FONT><FONT
color=black><SPAN style="COLOR: black"><BR
style="mso-special-character: line-break"><![if !supportLineBreakNewLine]><BR
style="mso-special-character: line-break"><![endif]></SPAN></FONT><FONT
color=black><SPAN
style="COLOR: black; mso-color-alt: windowtext"><o:p></o:p></SPAN></FONT></P>
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<P class=MsoNormal><B><FONT face=sans-serif color=black size=1><SPAN
style="FONT-WEIGHT: bold; FONT-SIZE: 7.5pt; COLOR: black; FONT-FAMILY: sans-serif">Pat
Flannery <pjfnefro@duke.edu></SPAN></FONT></B><FONT
color=black><SPAN style="COLOR: black"> <BR></SPAN></FONT><FONT
face=sans-serif color=black size=1><SPAN
style="FONT-SIZE: 7.5pt; COLOR: black; FONT-FAMILY: sans-serif">Sent by:
histonet-admin@lists.utsouthwestern.edu</SPAN></FONT><FONT
color=black><SPAN style="COLOR: black"> </SPAN></FONT><FONT
color=black><SPAN
style="COLOR: black; mso-color-alt: windowtext"><o:p></o:p></SPAN></FONT></P>
<P><FONT face=sans-serif color=black size=1><SPAN
style="FONT-SIZE: 7.5pt; COLOR: black; FONT-FAMILY: sans-serif">12/02/2003
03:00 PM</SPAN></FONT><FONT color=black><SPAN style="COLOR: black">
</SPAN></FONT><FONT color=black><SPAN
style="COLOR: black; mso-color-alt: windowtext"><o:p></o:p></SPAN></FONT></P></TD>
<TD
style="PADDING-RIGHT: 0.75pt; PADDING-LEFT: 0.75pt; PADDING-BOTTOM: 0.75pt; PADDING-TOP: 0.75pt"
vAlign=top>
<P class=MsoNormal><FONT face=Arial color=black size=1><SPAN
style="FONT-SIZE: 7.5pt; COLOR: black; FONT-FAMILY: Arial">
</SPAN></FONT><FONT color=black><SPAN
style="COLOR: black"><BR></SPAN></FONT><FONT face=sans-serif color=black
size=1><SPAN
style="FONT-SIZE: 7.5pt; COLOR: black; FONT-FAMILY: sans-serif">
To:
Histonet@lists.utsouthwestern.edu</SPAN></FONT><FONT
color=black><SPAN style="COLOR: black"> <BR></SPAN></FONT><FONT
face=sans-serif color=black size=1><SPAN
style="FONT-SIZE: 7.5pt; COLOR: black; FONT-FAMILY: sans-serif">
cc: </SPAN></FONT><FONT
color=black><SPAN style="COLOR: black"> <BR></SPAN></FONT><FONT
face=sans-serif color=black size=1><SPAN
style="FONT-SIZE: 7.5pt; COLOR: black; FONT-FAMILY: sans-serif">
Subject: [Histonet]
Citrate Buffer pH</SPAN></FONT><FONT color=black><SPAN
style="COLOR: black; mso-color-alt: windowtext"><o:p></o:p></SPAN></FONT></P></TD></TR></TBODY></TABLE>
<P class=MsoNormal
style="MARGIN-BOTTOM: 12pt; MARGIN-LEFT: 0.5in; MARGIN-RIGHT: 0in; mso-margin-top-alt: 0in"><FONT
face="Times New Roman" color=black size=3><SPAN
style="FONT-SIZE: 12pt; COLOR: black"><BR><BR><BR></SPAN></FONT><FONT
face="Courier New" color=black size=2><SPAN
style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: 'Courier New'">I have a
procedure for deparaffinizing (their word, not mine) tissue <BR>sections in
order to do immunostaining using the Vector MOM kit. It <BR>calls for
heating the section (briefly) in a microwave in 0.1M Citrate <BR>Buffer.
The procedure does not specify the pH of the buffer, just to
<BR>make it by mixing 0.1M NaCitrate and 0.1M citric acid. Does this
sound <BR>familiar to anyone out there?<BR><BR>-- <BR><BR>-Patrick J. (Pat)
Flannery<BR>Division of Nephrology (that's kidneys to you)<BR>Box 3014 (that's
NOT "PO" just "Box")<BR>Duke University Medical Center<BR>Durham, NC
27710<BR>E-mail: pjfnefro@duke.edu (preferred)<BR>
FLANN002@MC.DUKE.EDU (also works)<BR>Voice:
(919)660-6863<BR>Fax:
(919)684-2929<BR><BR><BR><BR><BR>_______________________________________________<BR>Histonet
mailing
list<BR>Histonet@lists.utsouthwestern.edu<BR>http://lists.utsouthwestern.edu/mailman/listinfo/histonet<BR
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