Re: "tunel" assay of Darcynkiewicz-Z

Andreas.Christaras (christar@uni-duesseldorf.de)
Fri, 1 Dec 1995 16:16:42 +0000

At 13:11 30.11.1995 -0500, you wrote:
>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)=20
>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.=20
>Zbigniew Darzynkiewicz
>
>
>
Date: 01.12.1995
Loc.: Duesseldorf, Germany
=20
Dear Dr. Darzynkiewicz,

I have a question concerning a commericially available tunel-assay kit (in
situ cell death detection, fluorescein) by Boehringer Mannheim Company,
Mannheim, Germany. This kit designed for FCM and immunofluorescence
microscopy uses diretly FITC-labelled dUTP in contradiction to digdUTP or
biotdUTP. Does the convalent linkage of FITC to dUTP provide any significant
advantage compared to the non-covalently linked two-step reagents?
Best regards,

Andreas Christaras

Andreas Christaras
Heinrich-Heine-University Duesseldorf
University Children=B4s Hospital
Laboratory of Experimental Hematology and Stem Cell Transplantation=
(KMT-Labor)
Building 14.82 (Gebaeude 14.82)
Moorenstrasse 5
40225 Duesseldorf
GERMANY
Voice : +49-211-311-6103
Fax : +49-211-311-6206
E-Mail : christar@uni-duesseldorf.de
Note : due to missing direct access to the internet at the moment, delays
in responding messages sent by E-mail can unfortunately occur.

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CD-ROM Vol 3 was produced by Monica M. Shively and other 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: (765)-494-0757; FAX(765) 494-0517; Web http://www.cyto.purdue.edu , EMAIL cdrom3@flowcyt.cyto.purdue.edu