Andrea,

I suspect that your problem lies with the Proteinase K.
This needs to be very carefully controlled. We had our most consistent
results by not using the proteolysis step and by adopting a standard
24hr fixation in formaldehyde, followed by 24hr in 80% ethanol prior to
wax processing. We used bowel mucosa as control tissue.

Regards

Bryan

>----------
>From: 	andreah@imclone.com[SMTP:andreah@imclone.com]
>Sent: 	October 20, 1998 5:21 PM
>To: 	HistoNet@Pathology.swmed.edu
>Subject: 	TUNEL CONTROLS
>
>Histonet,
>
>What suggestions could anyone give me for proper controlling of TUNEL
>assay done using the Boehringer Mannheim kit?  I know that it has been
>mentioned recently on Histonet that this assay needs to be highly
>controlled and since I seem to have WAY too many positive nuclei in my
>most recent experiment, I'd like some help ...
>
>Thank you in advance ...
>
>Andrea Hooper
>ANDREAH@IMCLONE.COM
>
>
>
>