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<DIV>Dear Tan</DIV>
<DIV>You have a lot of advaices in the wonderfull manual of Technical
Immunohistochemistry of Rodney Miller, go to <A
href="http://www.propathlab.com">www.propathlab.com</A> and download it free.Or
go to <A
href="http://hjaldanamarcos.bravepages.com/documentos.htm">http://hjaldanamarcos.bravepages.com/documentos.htm</A> and
donwload the documents named INMUNO1.... to INMUNO5.</DIV>
<DIV>By</DIV>
<DIV> </DIV></DIV>
<DIV>Dr. Hernán J. Aldana Marcos<BR>Facultad de Medicina. Universidad de
Morón<BR>Machado 914. B1708JPD. Buenos Aires. Argentina <BR>e-mail alternativo
<A href="mailto:hernanjavier@yahoo.com">hernanjavier@yahoo.com</A><BR>web: <A
href="http://hjaldanamarcos.bravepages.com">http://hjaldanamarcos.bravepages.com</A><BR><A
href="http://histologia.bigthicketdirectory.net/main.html">http://histologia.bigthicketdirectory.net/main.html</A><BR></DIV>
<BLOCKQUOTE
style="PADDING-RIGHT: 0px; PADDING-LEFT: 5px; MARGIN-LEFT: 5px; BORDER-LEFT: #000000 2px solid; MARGIN-RIGHT: 0px">
<DIV style="FONT: 10pt arial">----- Original Message ----- </DIV>
<DIV
style="BACKGROUND: #e4e4e4; FONT: 10pt arial; font-color: black"><B>From:</B>
<A title=MinHan.Tan@vai.org href="mailto:MinHan.Tan@vai.org">Tan, MinHan</A>
</DIV>
<DIV style="FONT: 10pt arial"><B>To:</B> <A
title=histonet@lists.utsouthwestern.edu
href="mailto:histonet@lists.utsouthwestern.edu">histonet@lists.utsouthwestern.edu</A>
</DIV>
<DIV style="FONT: 10pt arial"><B>Sent:</B> Wednesday, November 19, 2003 7:32
PM</DIV>
<DIV style="FONT: 10pt arial"><B>Subject:</B> [Histonet] Frozen section fixing
problems - but paraffin works!</DIV>
<DIV><BR></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2>Hi,</FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial size=2>I am working with
a new monoclonal antibody in parathyroid tissue, and I use the ABC-DAB
reageant system for immunohistochemistry.</FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial size=2>I am new to frozen
sections immunostaining - and I am encountering problems with frozen sections
- staining is very weak or absent, in comparison to paraffin embedded tissue,
which I have no problems with. </FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003></SPAN><SPAN
class=664142622-19112003><FONT face=Arial size=2>My protocol for frozen
sections: </FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial size=2>Unfixed frozen
slides with 5 micron sections thawed at room temperature for 5
minutes</FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial size=2>Placed in 70%
ethanol x 5 minutes.</FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial size=2>Washed in PBS x 5
minutes.</FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2>...</FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial size=2>followed with
standard protocol: hydrogen peroxide 0.3% x 30 min, donkey serum 5% x 30
min, primary antibody (mouse) 4 deg overnight, secondary (goat anti-mouse),
ABC, DAB.</FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003></SPAN><SPAN
class=664142622-19112003></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial size=2>Does anyone have
any advice?</FONT></SPAN></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial size=2>Thank
you!</FONT></SPAN></DIV><SPAN class=664142622-19112003>
<DIV><FONT face=Arial size=2></FONT><FONT face=Arial size=2></FONT><FONT
face=Arial size=2></FONT><FONT face=Arial size=2></FONT><FONT face=Arial
size=2></FONT><FONT face=Arial size=2></FONT><FONT face=Arial
size=2></FONT><FONT face=Arial size=2></FONT><BR><FONT face=Arial
size=2>Regards,</FONT></DIV>
<DIV><FONT face=Arial><FONT size=2><SPAN class=664142622-19112003>Min-Han
Tan</SPAN> </FONT></FONT></DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV></SPAN>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV>
<DIV><SPAN class=664142622-19112003><FONT face=Arial
size=2></FONT></SPAN> </DIV><FONT face=Arial>
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<STRONG></FONT>
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