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To: bionet-drosophila@net.bio.net
From: luschnig@pmgm2.stanford.edu (Stefan Luschnig)
Newsgroups: bionet.drosophila
Subject: Re: heat fixation
 dros@net.bio.netros@net.bio.net&replyto=3C7F688A.E63AA0BC@mail.uni-mainz.d
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Hi,

there is a protocol for heat fixation in the "Roberts " methods book 
("Drosophila: a practical approach") in the chapter "looking at 
embryos".
just briefly : you essentially collect embryos in a small steel mesh, 
throw them in boiling 0.1 % NaCl, 0.1% Triton-X 100 for 40 sec (in a 
scintillation vial), add more cold solution, and chill them on ice, 
before you devitellinize them in heptane/methanol as usual.  It 
seems to work well for certain proteins.

good luck,
Stefan

luschnig@cmgm.stanford.edu


-- 
___________________________________________________________
Stefan Luschnig
Department of Biochemistry/HHMI
279 Campus Drive, Beckman Center Rm. B453
Stanford University School of Medicine
Stanford, CA 94305
USA

lab phone	(650)-723-5872
Fax		(650)-723-6783
E-mail		luschnig@cmgm.stanford.edu
____________________________________________________________
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<html><head><style type="text/css"><!--
blockquote, dl, ul, ol, li { padding-top: 0 ; padding-bottom: 0 }
 --></style><title>Re: heat fixation
dros@net.bio.netros@net.bio.net&amp;repl</title></head><body>
<div><font face="Helvetica" size="+2" color="#000000">Hi,<br>
<br>
there is a protocol for heat fixation in the &quot;Roberts &quot;
methods book (&quot;Drosophila: a practical approach&quot;) in the
chapter &quot;looking at embryos&quot;.<br>
just briefly : you essentially collect embryos in a small steel mesh,
throw them in boiling 0.1 % NaCl, 0.1% Triton-X 100 for 40 sec (in a
scintillation vial), add more cold solution, and chill them on ice,
before you devitellinize them in heptane/methanol as usual.&nbsp; It&nbsp;
seems to work well for certain proteins.<br>
<br>
good luck,<br>
Stefan</font><br>
<font face="Helvetica" size="+2" color="#000000"></font></div>
<div><font face="Helvetica" size="+2"
color="#000000">luschnig@cmgm.stanford.edu</font><br>
<font face="Helvetica" size="+2" color="#000000"></font></div>
<div><br></div>
<x-sigsep><pre>-- 
</pre></x-sigsep>
<div>___________________________________________________________<br>
Stefan Luschnig<br>
Department of Biochemistry/HHMI<br>
279 Campus Drive, Beckman Center Rm. B453<br>
Stanford University School of Medicine<br>
Stanford, CA 94305<br>
USA<br>
<br>
lab phone<x-tab>&nbsp; </x-tab>(650)-723-5872<br>
Fax<x-tab>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</x-tab><x-tab>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</x-tab>(650)-723-6783<br>
E-mail<x-tab>&nbsp;&nbsp;&nbsp;
</x-tab><x-tab>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
</x-tab>luschnig@cmgm.stanford.edu<br>
____________________________________________________________</div>
</body>
</html>
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