From: ZBIGNIEW DARZYNKIEWICZ (darzynk@nymc.edu)
Date: Thu Nov 30 1995 - 13:11:50 EST
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
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