title: DRAQ5 labeling of nuclear DNA in live and fixed cells.
abstract: 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.
authors.affiliation: University of Wales College of Medicine, Heath Park, Cardiff, United Kingdom.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Smith
authors.forename: Marie
authors.initials: M
authors.lastname: Wiltshire
authors.forename: Rachel J
authors.initials: RJ
authors.lastname: Errington
mesh_terms: D000880:Anthraquinones; D002467:Cell Nucleus / Q000737:chemistry; D002477:Cells / Q000737:chemistry*; D004247:DNA / Q000032:analysis*; D003952:Diagnostic Imaging; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D008722:Methods
chemical_list: C520537:1,5-bis((2-(methylamino)ethyl)amino)-4,8-dihydroxyanthracene-9,10-dione; D000880:Anthraquinones; D005456:Fluorescent Dyes; D004247:DNA