Linda,
	Just finished Tunel on mouse brain. Cannot answer the question
re:fixation time. Used the ONCOR kit with a few modifications would happily
share protocol if you like. Very easy.
Cynthia Favara
NIAID/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
PH: 406-363-9317
FAX: 406-363-9286


-----Original Message-----
From: LINDA MARGRAF MD [mailto:LMARGRAF@childmed.dallas.tx.us]
Sent: Wednesday, March 24, 1999 12:13 PM
To: histonet@pathology.swmed.edu
Subject: TUNEL staining


Dear Histonetters:
I have been asked to help one of our researchers set up in-situ end labeling
or TUNEL staining for apoptosis in human autopsy brain. 
I have only a limited experience in doing the procedure and when we did it
in the past we used homemade reagents and skipped the kit.  My largest
concern is with the nature of the tissue we would be using tin this project.
The brains (mostly fetal and infant) are fixed for 1-2 weeks in 20% formalin
prior to processing and paraffin embedding. Does anyone know if this amount
of fixation will create too much DNA fragmentation to invalidate the
procedure. I would also love to hear from anyone doing this procedure about
kits or methods they have found helpful.
Thanks
Linda M
Histonet administrator
Associate Professor of Pathology
Southwestern Medical School
Dallas