Re: TUNEL

Eric Miller (miller@icrf.icnet.uk)
Fri, 12 Jan 1996 08:57:22 +0000 (GMT)

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
assays.

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@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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