Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1986-02-24
1989-03-14
Kepplinger, Esther M.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 7, 435 29, 435 91, 435810, 436 56, 436 63, 436 94, 436508, 436517, 436518, 436800, 935 77, 935108, C12Q 168
Patent
active
048123943
ABSTRACT:
A method is provided for simultaneously measuring total cellular DNA and incorporated nucleoside analog. The method entails altering the cellular DNA of cells grown in the presence of a nucleoside analog so that single stranded and double stranded portions are present. Separate stains are used against the two portions. An immunochemical stain is used against the single stranded portion to provide a measure of incorporated nucleoside analog, and a double strand DNA-specific stain is used against the double stranded portion to simultaneously provide a measure of total cellular DNA.
The method permits rapid flow cytometric analysis of cell populations, rapid identification of cycling and noncycling subpopulations, and determination of the efficacy of S phase cytotoxic anticancer agents.
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Darzykiewicz et al., Exp. Cell Res., vol. 90, (1975), pp. 411-428.
Gratzner et al., J. Histochem. Cytochem., vol. 24, (1976), pp. 34-39.
Inman, Methods in Enz., vol. 29, (1974), pp. 451-458.
Darzykiewicz et al., J. Histochem. Cytochem., vol. 24, (1976), pp. 49-58.
Darzynkiewicz et al., J. Histochem. Cytochem., vol. 27, (1979), pp. 478-485.
Moran et al., J. Histochem. Cytochem., vol. 33, (1985), pp. 821-827.
Gratzner et al., Exp. Cell Res., vol. 95, (1975), pp. 88-94.
Gratzner et al., Cytometry, vol. 1, (1981), pp. 385-389.
Dolbeare Frank A.
Gray Joe W.
Kepplinger Esther M.
Macevicz Stephen C.
Sartorio Henry P.
University of California
Wieder Stephen C.
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