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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