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Title: Flow cytometric measurement of total DNA content and incorporated promodeoxyuridine

Abstract

A procedure was developed for simultaneous flow cytometric measurement of cellular DNA content and amount of BrdUrd incorporated into cellular DNA. Propidium iodide was used as a fluorescent probe for total cellular DNA and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescein-labeled goat anti-mouse antibody was used to fluorescently label the bound anti-BrdUrd probe. Bivariate DNA/BrdUrd distributions measured for Chinese hamster ovary cells labeled for 30 min with BrdUrd clearly show the G/sub 1/- and G/sub 2/M-phase cells to have low BrdUrd-linked fluorescence and the S-phase cells to have high BrdUrd-linked fluorescence. Cell cycle traverse rates were estimated for Chinese hamster ovary cells from bivariate distributions measured for samples taken periodically after pulse labeling with BrdUrd. Bivariate DNA/BrdUrd distributions were also applied in the analysis of the response of C3H murine bone marrow cells to treatment in vivo with 1-..beta..-D-arabinofuranosylcytosine (araC). Bivariate distributions were measured for bone marrow cells taken from mice that were pulse labeled with BrdUrd at various times after treatment with araC. The resulting DNA/BrdUrd sequences show the kinetics of recovery from araC and allow discrimination of the araC sterilized cells.

Authors:
; ; ;
Publication Date:
Research Org.:
Lawrence Livermore National Lab., CA
OSTI Identifier:
6008004
DOE Contract Number:  
W-7405-ENG-48
Resource Type:
Journal Article
Journal Name:
Proc. Natl. Acad. Sci. U.S.A.; (United States)
Additional Journal Information:
Journal Volume: 80
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ANIMAL CELLS; ANTIBODIES; ANTIMETABOLITES; AZINES; BROMOURACILS; DRUGS; HETEROCYCLIC COMPOUNDS; HYDROXY COMPOUNDS; NUCLEIC ACIDS; NUCLEOSIDES; NUCLEOTIDES; ORGANIC BROMINE COMPOUNDS; ORGANIC COMPOUNDS; ORGANIC HALOGEN COMPOUNDS; ORGANIC NIT; 550200* - Biochemistry

Citation Formats

Dolbeare, F, Gratzner, H, Pallavicini, M G, and Gray, J W. Flow cytometric measurement of total DNA content and incorporated promodeoxyuridine. United States: N. p., 1983. Web. doi:10.1073/pnas.80.18.5573.
Dolbeare, F, Gratzner, H, Pallavicini, M G, & Gray, J W. Flow cytometric measurement of total DNA content and incorporated promodeoxyuridine. United States. https://doi.org/10.1073/pnas.80.18.5573
Dolbeare, F, Gratzner, H, Pallavicini, M G, and Gray, J W. Thu . "Flow cytometric measurement of total DNA content and incorporated promodeoxyuridine". United States. https://doi.org/10.1073/pnas.80.18.5573.
@article{osti_6008004,
title = {Flow cytometric measurement of total DNA content and incorporated promodeoxyuridine},
author = {Dolbeare, F and Gratzner, H and Pallavicini, M G and Gray, J W},
abstractNote = {A procedure was developed for simultaneous flow cytometric measurement of cellular DNA content and amount of BrdUrd incorporated into cellular DNA. Propidium iodide was used as a fluorescent probe for total cellular DNA and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescein-labeled goat anti-mouse antibody was used to fluorescently label the bound anti-BrdUrd probe. Bivariate DNA/BrdUrd distributions measured for Chinese hamster ovary cells labeled for 30 min with BrdUrd clearly show the G/sub 1/- and G/sub 2/M-phase cells to have low BrdUrd-linked fluorescence and the S-phase cells to have high BrdUrd-linked fluorescence. Cell cycle traverse rates were estimated for Chinese hamster ovary cells from bivariate distributions measured for samples taken periodically after pulse labeling with BrdUrd. Bivariate DNA/BrdUrd distributions were also applied in the analysis of the response of C3H murine bone marrow cells to treatment in vivo with 1-..beta..-D-arabinofuranosylcytosine (araC). Bivariate distributions were measured for bone marrow cells taken from mice that were pulse labeled with BrdUrd at various times after treatment with araC. The resulting DNA/BrdUrd sequences show the kinetics of recovery from araC and allow discrimination of the araC sterilized cells.},
doi = {10.1073/pnas.80.18.5573},
url = {https://www.osti.gov/biblio/6008004}, journal = {Proc. Natl. Acad. Sci. U.S.A.; (United States)},
number = ,
volume = 80,
place = {United States},
year = {1983},
month = {9}
}