Eric Miller miller at icrf.icnet.uk
Fri Jan 12 03:57:22 EST 1996

Can't help you on the nuclease issue: what we use is the cell line COLO 205.
This line is exceptionally susceptible to apoptosis, and after 2-3 days 
of serum deprivation will give you 40-50% apoptotic cells +ve for TUNEL 

	Eric P Miller
	Edinburgh Medical Oncology Unit

"I am Pentium of Borg. Division is futile......you will be approximated."
Star Trek - Generations. Two Captains, one density.

On Thu, 11 Jan 1996, Dan Smith wrote:

> I am forwarding a request from a coworker regarding getting a GOOD positive 
> control to work in our TUNEL/apoptosis assay.    Which nuclease have you 
> used (company, ph#, etc) with the best results.
> Thanks again!
> Dan Smith
> San Diego
> e-mail: djs at allp.com (or through the users group)
>  ----------
> We have been unable to get the positive control to work in her TUNEL 
> protocols. The control involves treating the permeabilized cells with a 
> nuclease to nick the DNA and render it TUNEL positive.  It must be a problem 
> with the enzyme itself since we are registering what we interpret as TUNEL 
> positive cells in the treated samples. Could you submit a question to the 
> internet group concerning what nuclease and vendor might help us solve this 
> problem. We could just start ordering different ones from different places 
> but this may give us a more reasonable approach. Long live the NET!

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