7-Aminoactinomycin D

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
1. Solution preparation
1.1 Preparation of stock solution
Solvent: DMSO
Concentration: 1-10 mM.
Storage: Store at -20°C or -80°C in dark after aliquoting. Avoid repeated freezing and thawing.
1.2 Preparation of working solution
Dilute to 0.5-5 µM with PBS or serum-free medium (optimized according to the experiment).
Note: The working solution should be prepared and used immediately. Keep it away from light.

2. Cell staining
2.1 Fix cells.
2.2 Pellet and resuspend cells.
2.3 Add 7-Aminoactinomycin D (7-AAD) staining solution (0.5 to 5 µM).
2.4 Incubate for 15 to 60 minutes.
2.5 Measure fluorescence intensity (Ex/Em = 549/648 nm).

7-Aminoactinomycin D (7-AAD) is a DNA dye that distinguishes viable, apoptotic, and late apoptotic/dead cells in flow cytometry^[1].
7-Aminoactinomycin D is frequently used to stain and exclude dead cells in flow cytometry at low concentrations (0.5-5 μg/mL). At higher concentrations (10-20 μg/mL), 7-Aminoactinomycin D has also been used to distinguish between viable cells (7-AAD_negative) and apoptotic cells (7-AAD_dim) or dead cells (7-AAD_bright) using the fact that permeability of the cell membrane, and hence fluorescence intensity, is low in early apoptotic cells and high in late apoptotic and dead cells^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.