7-aminoactinomycin (7-AAD) is a fluorescent dye that has a high affinity for GC-rich regions of double stranded DNA. 7-AAD is not membrane permeable and therefore requires the compromised membranes of dead cells to bind to DNA. Upon excitation with an appropriate light source, dead cells exhibit higher fluorescence intensity than live cells, thereby enabling clear discrimination of these populations by flow cytometry. Dead cells can then be excluded from further analysis and also sorted from cell populations of interest. The spectral properties (Excitation 546 nm/ Emission 647 nm) enable use in both single color and multi color flow experiments. The convenient dropper bottle format removes the manual weighing, pipetting and dilution steps required by other commonly available 7-AAD products.
- Store at room temperature
- Guaranteed until date of expiry. Please see product label.
- For research purposes only
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Applications of READIDROP™ 7-AAD
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Instructions For Use
- Caution: Wear gloves, safety glasses/face protection, and protective clothing to avoid direct contact of ReadiDrop solutions with skin and eyes.
1. Resuspend 1 x 106 cells in 500 μl flow cytometry buffer (1x phosphate buffered saline with 3% BSA). The cell concentration is now 2 x 106/ml.
2. Add one to two drop of ReadiDrop 7-AAD to the cell suspension.
3. Vortex briefly.
4. Incubate for 1-10 min at room temperature.
Note: Adding one drop to a 500 µl cell suspension produces a final dye concentration of 1 μg/ml. This is typically sufficient to stain 1 x 106 cells. If one drop is not sufficient for your cell type, adding a second drop may improve staining efficacy. When working with a larger volume of cells, scale ReadiDrop solution amount accordingly.
5. Analyze cells by flow cytometry.
- Detect 7-AAD signal in FL3 on Bio-Rad's S3e Cell Sorter. Otherwise, select the most appropriate FL channel on your particular instrument.
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How to Use the Spectraviewer
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Start by selecting the application you are interested in, with the option to select an instrument from the drop down menu or create a customized instrument
Select the fluorophores or fluorescent proteins you want to include in your panel to check compatibility
Select the lasers and filters you wish to include
Select combined or multi-laser view to visualize the spectra