Description
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.
Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Development References (1)
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Schmid I, Krall WJ, Uittenbogaart CH, Braun J, Giorgi JV. Dead cell discrimination with 7-amino-actinomycin D in combination with dual color immunofluorescence in single laser flow cytometry. Cytometry. 1992; 13(2):204-208. (Methodology: Flow cytometry). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.