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
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