DRAQ5 Labeling of Nuclear DNA in Live and Fixed Cells
This unit describes the use of a novel DNA-detecting far-red-fluorescing dye, DRAQ5, a modified anthraquinone, which has a unique combination of properties exploitable by cytometry. These include a high capacity to permeate the cell membrane, a high DNA binding affinity and selectivity, a fluorescence emission spectrum beyond that of fluorescein, phycoerythrin, Texas Red, Cy3, and EGFP, and excitation characteristics separate from those of propidium iodide. In this unit, methods are presented for preparation and analysis of both live and fixed cells stained with DRAQ5. While the focus is on flow cytometric assays, typical imaging applications are also indicated because the staining protocols share the same essential features.
Keywords: anthraquinone; intercalation; DNA staining
Basic Protocol: Preparation and DRAQ5 Staining of Live Cells for Analysis Using Flow Cytometry or Imaging
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This material safety data sheet provides health and safety information for DRAQ5.
The Flow-Cytometry Laboratory of the Department of Clinical Research, University of Bern, has provided a protocol for immunophenotyping that includes DRAQ5 staining.
These sites provide examples of fix-and-stain protocols using DRAQ5.
Research supported by the UK Research Councils' Basic Technology Research Programme Grant GR/S23483 and UK Biotechnology and Biological Sciences Research Council Grant SBRI19666.