Hello If you use FITC-labeled dUTP in your TUNEL, combining TUNEL with PI might give you problems. The reason is that propidium iodide might quench the fluorescence of TdT-incorporated FITC-labeled dUTP in apoptotic cells. Look up the following paper: Trond Stokke et al. Cytometry 33: 428-434 (1998). Unni Haddeland, PhD Research Laboratory University College of Akershus Ringstabekkvn 105 1356 Bekkestua Norway Den 9 Dec 99 klokken 23:55, skrev Nan : > Date: Thu, 09 Dec 1999 23:55:05 -0600 > From: Nan <Nan.Lin-2@tc.umn.edu> > Reply-to: Nan Lin <Nan.Lin-2@tc.umn.edu> > To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> > Subject: taxol and TUNEL > > Hello all: > I'm trying to measure apoptosis induced by taxol (paclitaxel) and some > other chemicals in MCF-7 cells by 2-color TUNEL, in order to correlate > apoptosis and cell cycle. I used PI to stain the DNA. I have tried > several doses and treatment time of taxol, but didn't get any positive > TUNEL labelling. My other chemicals all showed positive TUNEL labelling, > and taxol did show G2/M arrest and later sub-G1 peak as reported by > others and as indicated by FL2. But NO TUNEL positive population! > > I noticed some discussion of "taxol and TUNEL incompatibility" in the > mailing list a couple of years ago. I wonder if any one had worked with > TUNEL and taxol and would help me out? Taxol is supposed to be my > positive control. Others prepared slides and reported TUNEL positive > results with the same doses and treatment time as I did. Is it my > method, or is the TUNEL doesn't work well with taxol? > Any suggestions on 2-color TUNEL is appreciated! > > thanks > Nan Lin > >
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