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Effect of 5‐fluoro‐2′‐deoxyuridine (FdUrd) on 5‐bromo‐2′‐deoxyuridine (BrdUrd) incorporation into DNA, measured with a monoclonal BrdUrd antibody and by the BrdUrd/hoechst quenching effect

Effect of 5‐fluoro‐2′‐deoxyuridine (FdUrd) on 5‐bromo‐2′‐deoxyuridine (BrdUrd) incorporation into... The purpose of this study was to improve the application of bromodeoxyuridine (BrdUrd) for the flow cytometric analysis of cell kinetics. In order to obtain a quantitative measure of the DNA synthesis rate (or the number of divided cells). BrdUrd should replace thymidine (dThd) completely in the newly synthesized DNA strands. The de novo synthesis of dThd monophosphate competing with BrdUrd incorporation was stopped by fluorodeoxyuridine (FdUrd). Cells of a human leukemic cell line (REH) were exposed O BrdUrd for either 20 min, 8h, or 24h. Bromodeoxyuridine incorporation was determined by a monoclonal antibody as well as by the BrdUrd/Hoechst (H) technique. Counter‐staining of the DNA was performed with propidium iodide or ethidium bromide. DNA fluorescences were measured in both techniques with a two‐parameter flow cytometer, the his‐tograms being analyzed by computer. It was found that FdUrd is required in the BrdUrd/H technique for replacement of dThd at low BrdUrd concentrations and long incubation times. With short incubation periods, as used for detection by the monoclonal anti‐BrdUrd antibody, FdUrd increases the incorporated BrdUrd amount when BrdUrd concentrations of 10 μU or less are applied. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Cytometry Part A Wiley

Effect of 5‐fluoro‐2′‐deoxyuridine (FdUrd) on 5‐bromo‐2′‐deoxyuridine (BrdUrd) incorporation into DNA, measured with a monoclonal BrdUrd antibody and by the BrdUrd/hoechst quenching effect

Ellwart, Joachim; Dörmer, Peter
Cytometry Part A , Volume 6 (6) – Nov 1, 1985
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References (40)

Publisher
Wiley
Copyright
Copyright © 1985 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1552-4922
eISSN
1552-4930
DOI
10.1002/cyto.990060605
pmid
2415309
Publisher site
See Article on Publisher Site

Abstract

The purpose of this study was to improve the application of bromodeoxyuridine (BrdUrd) for the flow cytometric analysis of cell kinetics. In order to obtain a quantitative measure of the DNA synthesis rate (or the number of divided cells). BrdUrd should replace thymidine (dThd) completely in the newly synthesized DNA strands. The de novo synthesis of dThd monophosphate competing with BrdUrd incorporation was stopped by fluorodeoxyuridine (FdUrd). Cells of a human leukemic cell line (REH) were exposed O BrdUrd for either 20 min, 8h, or 24h. Bromodeoxyuridine incorporation was determined by a monoclonal antibody as well as by the BrdUrd/Hoechst (H) technique. Counter‐staining of the DNA was performed with propidium iodide or ethidium bromide. DNA fluorescences were measured in both techniques with a two‐parameter flow cytometer, the his‐tograms being analyzed by computer. It was found that FdUrd is required in the BrdUrd/H technique for replacement of dThd at low BrdUrd concentrations and long incubation times. With short incubation periods, as used for detection by the monoclonal anti‐BrdUrd antibody, FdUrd increases the incorporated BrdUrd amount when BrdUrd concentrations of 10 μU or less are applied.

Journal

Cytometry Part A – Wiley

Published: Nov 1, 1985

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