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Live or Dead™ Fixable Dead Cell Staining Kit *IR Fluorescence*
Example protocol
AT A GLANCE
Prepare samples in HHBS buffer (0.5 mL/assay).
Wash cells and replace with fresh HHBS buffer.
Add Stain It™ IR fluorescence to the cell suspension.
- Stain the cells at room temperature or 37°C for 20 - 60 minutes
Wash the cells.
Fix the cells (optional).
Examine the sample with a flow cytometer using an 808 nm laser with an 885/40 nm or IR channel emission filter.
Before starting the experiment, thaw all the components at room temperature.
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Add 200 µL of DMSO (Component B) to the vial of Stain It™ IR fluorescence (Component A) to make a 500X Stain It™ IR fluorescence stock solution.
SAMPLE EXPERIMENTAL PROTOCOL
Prepare cells using 1X Hanks and 20 mM Hepes buffer (HHBS) or a sodium azide-free and serum/protein-free buffer of your choice.
Wash the cells once with HHBS or the azide- and serum/protein-free buffer of your choice.
Resuspend the cells at 5 - 10 × 106/mL in HHBS or in the azide- and serum/protein-free buffer of your choice.
Add 1 µL of 500X Stain It™ IR fluorescence stock solution to 0.5 mL of cells/assay and mix well.
Incubate at room temperature or 37°C, 5% CO2 incubator for 20 - 60 minutes, protected from light.
Note: The optimal stain concentrations and incubation time should be experimentally determined for different cell lines.
Wash the cells twice and resuspend cells with HHBS or a buffer of your choice.
Fix cells as desired (optional).
Analyze cells with a flow cytometer using the appropriate excitation/emission specifications found in the 'Key Parameters' section.
Safety data sheet