"tunel" assay
ZBIGNIEW DARZYNKIEWICZ (darzynk@nymc.edu)
Thu, 30 Nov 1995 13:11:50 -0500
Dr. Petrie inquires about the commercial kits to label DNA strand
breaks for analysis of apoptosis. We have recently improved the assay
using BrdUTP as the labeled deoxynucleotide and then applying BrdUrd
MoAb-FITC conjugate (from Becton Dickinson) to detect the
incorporated BrdUTP. The method has several advantages. The cost of
BrdUTP (Sigma)is 1000X (yes thousand) lower than of the digoxygenin
or biotin labeled nucleotides. Furthermore, the method is twice more
sensitive that the ones utlizing digdUTP or biotdUTP (indirect, two
step procedures), and eight times more sensitive than the single step
labeling. The method also is less complex (fewer centrifugations)
then the two step procedures. The high sensitivity may be due to the
fact that BrdUTP has no bulky conjugates and thus may be incorporated
at better efficiency than e.g. digdUTP. The paper will appear in Dec
issue of Cell Proliferation and another in Jan issue of Exp. Cell
Res. I will be glad to send the protocol for the researchers who may
be interested in this approach. Please fax me (914-347-2804) your fax
number.
Zbigniew Darzynkiewicz
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
