The LIVE/DEAD® Fixable Blue Dead Cell Stain Kit is used to determine the viability of cells prior to the fixation and permeabilization required for intracellular antibody staining or prior to elimination of biohazardous materials using formaldehyde fixation. This kit has been optimized and validated for use with a violet laser flow cytometer.
• Stable—dyes are freeze dried in separate vials to maintain stability
• Robust—staining pattern is the same before and after fixation
• Bright signal—allows for easy distinction between live/dead cells in single channel
Unlike products that are sold in solution, the LIVE/DEAD® Fixable Blue Stain has been conveniently packaged in five 40-test vials to help ensure the stability and performance of the dye over time. Amine reactive dyes in solution will lose their effectiveness over a short period of time, therefore it is recommended to completely use the vial once rehydrated. If this is not possible, aliquot the vials in small volumes and store at -80°C, avoiding freeze-thaw cycles.
Dead cell discriminator stains can lose sensitivity after treatment with fixatives such as formaldehyde or ethanol-based fixation methods required for intracellular phosphorylation studies. The LIVE/DEAD® Fixable Blue Stain is an amine reactive dye that binds covalently to intracellular and extracellular amines, and the staining pattern is preserved following formaldehyde fixation.
The LIVE/DEAD® Fixable Blue Stain was selected based on its fluorescent properties to provide a bright signal when excited with a UV laser. The blue-fluorescent reactive dye has an excitation maximum of ~350 nm, making it ideal for use with an UV laser, and an emission of ~450 nm. By switching viability staining from a channel off of the blue 488 nm laser to the UV laser, you will free up a channel on your flow cytometer for other reagents that are difficult to find in colors other than FITC or PE. By using the UV laser, there is no spectral overlap issues with other common dyes, thus eliminating the need to compensate the viability stains from other stains in your panel. Since live and dead cells can be discriminated using a single dye and a single channel of a flow cytometer, it is an ideal choice for multi-color experiments.
How It Works
In cells with compromised membranes, the dye reacts with free amines both in the cell interior and on the cell surface, yielding intense fluorescent staining. In viable cells, the dye's reactivity is restricted to the cell-surface amines, resulting in less intense fluorescence. The difference in intensity is typically greater than 50-fold between live and dead cells, allowing for easy discrimination.
LIVE/DEAD® Fixable Dead Cell Stains are available in a wide variety of colors to meet your multi-color panel needs. See the LIVE/DEAD® Fixable Dead Cell Stains page
for more information.