eBioscience™ Fixable Viability Dye eFluor™ 450

The eBioscience Fixable Viability Dye is an amine-reactive dye and covalently binds to primary amines, such as those present on proteins. Amine-reactive dyes cross the cell membrane of dead cells, and react with free amines in the cytoplasm. Live cells exclude the dye because their cell membranes are intact. Samples that are stained with this viability dye show a bright and dim population. The very bright cells are dead, while the dimmer population represents live cells. This is because the dye can bind to all amines (extracellular and intracellular) in dead cells, whereas it can only stain surface amines in viable cells.

Yes, eFluor 450-conjugated antibodies are compatible with eBioscience Annexin V Apoptosis Detection kits.

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Discrimination between viable and non-viable cells can be carried out with the use of the 7-AAD Viability Dye (Cat. No. 00-6993-50) or Propidium Iodide (PI) Staining Solution (Cat. No. 00-6990-50). The 7-AAD or PI will mark the non-viable cells by binding to the nuclei of those cells. The nucleic acid of viable cells will not be accessible to the dye and will not be stained. When analyzing the data collected, gate out all cells stained with the viability dye. When staining for intracellular proteins, use the Fixable Viability Dye eFluor 455UV, 450, 520, 660, or 780.

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Yes, a Fixable Viability Dye can be added to samples after BrdU labeling and before fixation with the BrdU Staining Buffer. See the protocol for additional information.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.