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Just thought I'd contribute my 50 cents worth (inflation!) about using
camptothecin on HL60 cells as a positive control for TUNEL. As Dr.
Darzynkiewicz has said, it is very reliable. This has become our
apoptosis gold standard. I cannot overemphasize the ease in
generating this control. (Unless, of course, you do not have HL60
in-house; then, I suppose ATCC is the next stop...).
Hi!
Just thought I'd contribute my 50 cents worth (inflation!) about the use of
camptothecin ( buy it from Sigma; start with 0.15uM for 4 hours) on HL60
(maintain the cells as described in Cancer Research, vol. 53, pages 1945-1951,
1993, by Gorczyca, Gong, and Darzynkiewicz) as a positive control for apoptosis.
As Dr. Darzynkiewicz has said, this is a very reliable control!!! The ease in
generating this control is amazing. It has become our gold standard for
apoptosis. Please consider using this!
Linda Weaver
______________________________ Reply Separator _________________________________
Subject: Re: DNAse and TdT apoptosis assay
Author: owner-cyto-sendout@flowcyt.cyto.purdue.edu at INTERNET1
Date: 7/17/96 4:39 AM
>We have a client who wants to generate a postive control for the TdT
>apoptosis assay by treating normal cells with DNAse. I have a few questions
>for anyone who has done this.
Jeff,
Why not use a drug like camptothecin or etoposide?
If you really need to use DNase, you should try to find some of the in vivo
footprinting studies (sorry, I can't remember the citations)
--Russ Boggs
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