Ghost Dye™ Red 710 Fixable Viability Dye

Product Usage Information

1. Prepare the following reagents with reverse osmosis deionized (RODI) or equivalent grade water:a. 1X PBS (azide- and protein/serum-free)b. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.2. Remove Ghost Dye™ from -20°C and bring to room temperature.3. Collect cells by centrifugation and aspirate supernatant.4. Wash cells by centrifugation in excess 1X PBS. Repeat if necessary.5. Resuspend cells to a concentration of 1-10 x 106/mL in 1X PBS.6. Centrifuge the Ghost Dye™ before opening then add 1 uL for each 1 mL of cell suspension and vortex immediately.7. Incubate for 30 minutes at 4°C protected from light.8. Wash by centrifugation in excess incubation buffer. Discard supernatant. Repeat.9. Cells can then be fixed, permeabilized, and immunostained based upon experimental design and recommended protocols.10. Exclude cells with high Ghost Dye™ fluorescence from analysis. These were non-viable cells at the time of fixation. See details below for excitation and emission specifications. Ghost Dye™ Red 710 Fixable Viability Dye excited by the red (633-647 nm) laser line and has a peak emission of 710 nm that can be detected using the recommended 710/50 band pass filter commonly used for detection of Alexa Fluor® 700.