[Histonet] (no subject)
mab70 <@t> medschl.cam.ac.uk
Wed Mar 23 02:44:21 CST 2005
You should air dry your sections for about 1 hour. The OCT will wash off in
the wash buffers. Whether you need to do an avidin/biotin block will depend
upon the tissue, but you should try with and without to test this. You may
need to experiment with fixation, look at the datasheet for the antibody and
follow the recommendations. You may need to use paraformaldehyde fixation or
even plp for which you should be able to find the formula in a good
textbook. I use Tris buffered saline (TBS) for washing, others prefer
phosphate buffered saline (PBS). You will find a good protocol in the pack
insert in any of Vector's ABC kits, which are excellent. these pack inserts
also have a trouble shooting guide to help you.
I hope this helps. If you want a copy of my protocol, please contact me.
University of Cambridge
From: Margaryan, Naira [mailto:NMargaryan <@t> childrensmemorial.org]
Sent: Tuesday, March 22, 2005 8:19 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] (no subject)
I am going to do Immunostaining of Frozen Sections .
I have some questions:
1. What to do with slides after remove them from --80 o C freezer?
2. How long the slides have to come to room temperature?
3. How the OCT compound has to be dissolved?
4. What is general protocol for frozen sections?
5. Am I have to apply Avidin/biotin?
My appreciation for any suggestion,
Naira V. Margaryan, Ph.D, D.V.M.
Research Associate II
Children's Memorial Research Center
2300 Children's Plaza, Box 222
Chicago, IL 60614-3394
nmargaryan <@t> childrensmemorial.org
<mailto:nmargaryan <@t> childrensmemorial.org>
For Express Mail:
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