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.
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light.Do not freeze.
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/pharmingen/protocols for technical protocols.
