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<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><HR><STRONG></FONT>
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