Application

Fluorescent DNA stain. A featured product for Apoptosis research.

Biochem/physiol Actions

7-amino-actinomycin D (7-AAD) is a DNA dye. It can be used as a replacement for propidium iodide (PI) for the discrimination of dead cells. This dye is ideal for dead cell discrimination in long-lasting experiments, as it is efficiently exempted by intact cells and has a high DNA binding constant.

7-AAD is used in flow cytometry analysis of viable cells. 7-ADD is a fluorescent indicator that undergoes a spectral shift upon association with DNA. Su, X., et al. used FITC and PE-conjugated antibodies to stain cell surface markers. After cell-surface marker staining, cells were further stained with 10 µg/ml of 7-AAD in PBS on ice for 30 minutes. After washing with PBS twice, the cells were fixed in 1% paraformaldehyde supplemented with 50 µg/ml actinomycin D. Non-apoptotic cells were 7-AAD negative. 7-AAD like its parent molecule, Actinomycin D, is a DNA-intercalator with growth-inhibitory properties.

General description

Free form: Excitation: 503 nm (0.01 M phosphate buffer, pH 7.0 containing 0.1 mM EDTA); 550 nmEmission: 675 nm (0.01 M phosphate buffer, pH 7.0 containing 0.1 mM EDTA); 672 nmDNA Complex: Excitation: 543 nm; 555 nmEmission: 655 nm; 665 nm

Reconstitution

Solution of 1 mg/ml in DMSO:H2O (1:1) is stable for at least 2 years at −20 °C without loss of activity. According to Su et al., the provided solution should be diluted to 10 µg/ml (1:100) working concentration in PBS and is therefore sufficient for 100 stain reactions.