Invitrogen™ eBioscience™ Fixable Viability Dye eFluor™ 450

Catalog No.	Quantity
50-112-8817	100 Tests
50-169-62	500 Tests

Catalog No. 50-112-8817 Supplier Invitrogen™ Supplier No. 65086314

Description

A viability dye that can be used to irreversibly label dead cells prior to cryopreservation, fixation, and/or permeabilization procedures.

• Unlike 7-AAD and propidium iodide, cells labeled with Fixable Viability Dyes can be washed, fixed, permabilized, and stained for intracellular antigens without any loss of staining intensity of the dead cells
• Thus, using these dyes allows dead cells to be excluded from analysis when intracellular targets are being studied
• Fixable Viability Dye eFluor™ 450 can be excited by the violet (405 nm) laser line and has a peak excitation of 450 nm that can be detected using a 450/50 band pass filter (equivalent to eFluor™ 450 or Pacific Blue™)
• For compensation, it is recommended to use a sample of the cells of interest stained with the Fixable Viability Dye
• If the percentage of dead cells is expected to be less than 5%, then it is recommended to take a small aliquot of cells and heat them at 65°C for 1 minute then immediately place on ice for 1 minute
• After this treatment, the heat-killed cells can be combined 1:1 with live cells and then stained with the Fixable Viability Dye
• Fixable Viability Dye eFluor™ 450 is supplied as a pre-diluted solution prepared in high-quality, anhydrous DMSO
• It should be protected from light and moisture
• Store at -70°C with dessicant, it may be freeze-thawed up to 20 times
• Allow vial to equilibrate to room temperature before opening

Specifications

• Color: Blue
• Detection Method: Fluorescence
• For Use With (Application): Viability Assay
• For Use With (Equipment): Flow Cytometer
• Product Type: Dye
• Excitation Wavelength Range: 405/450
• Product Line: eBioscience
• Quantity: 100 Tests

Frequently Asked Questions (FAQs)

Why is there a slight increase in fluorescence for viable cells when using eBioscience Fixable Viability Dye (Cat. No. 65-0863-18, 65-0863-14)?

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.

Can I use eFluor 450-conjugated antibodies with eBioscience Annexin V Apoptosis Detection kits?

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

How do I discriminate between live cells and dead cells when doing a flow cytometry analysis?

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.

Can Fixable Viability Dyes be used with the eBioscience BrdU Staining kits for Flow Cytometry?

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.

For Research Use Only. Not for use in diagnostic procedures.