Cell‐Cycle Analysis Using A Monoclonal Antibody to Brdurd
- 1 July 1984
- journal article
- research article
- Published by Wiley in Cell Proliferation
- Vol. 17 (4) , 427-436
- https://doi.org/10.1111/j.1365-2184.1984.tb00601.x
Abstract
The flow cytometric measurement of DNA distributions of cells has many applications in biomedical research. Phase fractions estimated (calculated) from such distributions are used to study the growth characteristics of various types of cells, particularly when the cells have been exposed to perturbing agents such as chemotherapeutic drugs. For more than 10 years many methods for resolving DNA distributions into the three cell subpopulations (G1, S and G2, + M) have been reported in the literature. A new method of analysis utilizing a monoclonal antibody to bromodeoxyuridine (BrdUrd) has been developed (Gratzner, 1982; Dolbeare et al., 1983) which makes it possible in most cases to accurately determine phase fractions without resorting to mathematical models. the procedure involves the incorporation of BrdUrd by growing (DNA synthesizing) S phase cells, labelling the BrdUrd with a fluorescent monoclonal antibody, and the bivariate measurement of the antibody and of total DNA content, the latter through propidium‐iodide staining. the resulting bivariate distributions clearly and simply resolve the three subpopulations. This paper describes the method and illustrates its use in the analysis of various fractions of elutriated exponentially growing Chinese hamster ovary (CHO) cells.This publication has 6 references indexed in Scilit:
- Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine.Proceedings of the National Academy of Sciences, 1983
- The analysis and interpretation of DNA distributions measured by flow cytometryCytometry, 1982
- Monoclonal Antibody to 5-Bromo- and 5-Iododeoxyuridine: A New Reagent for Detection of DNA ReplicationScience, 1982
- Rates of incorporation of radioactive molecules during the cell cycleJournal of Cellular Physiology, 1981
- MATHEMATICAL ANALYSIS OF DNA DISTRIBUTIONS DERIVED FROM FLOW MICROFLUOROMETRYThe Journal of cell biology, 1974
- Cell Microfluorometry: A Method for Rapid Fluorescence MeasurementScience, 1969