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7-Amino-Actinomycin D (7-AAD) is a convenient, ready-to-use nucleic acid dye that can be used in place of propidium iodide (PI) for the exclusion of nonviable cells in flow cytometric assays. The advantage of 7-AAD over PI is the ability to be used in conjunction with phycoerythrin (PE)- and fluorescein isothiocyanate (FITC)-labelled monoclonal antibodies in 2-color analysis, with minimal spectral overlap between 7-AAD, PE and FITC fluorescence emissions. The 7-AAD fluorescence is detected in the far red range of the spectrum (650 nm long-pass filter). This reagent is used as a viability probe for methods of dead cell exclusion, based on light scatter and uptake of 7-AAD as detected in FL3. This reagent does not require dilution. Suggested quantity to use: 5 µl (0.25 µg)/test (1x10^6 cells) and incubate for 10 minutes before analysis.
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