Cell Explorer™ Live Cell Tracking Kit *Red Fluorescence*

Protocol summary

1. Prepare samples
2. Add 10X Track It™ Red working solution (10 µL/well)
3. Incubate the cells at 37°C for 15 to 60 minutes
4. Wash the cells
5. Examine the specimen under fluorescence microscope with Texas Red filter or flow cytometer with 610/20 nm filter (PE-Texas Red channel)

Important notes
Thaw all the components at room temperature. Centrifuge the Component A brieftly before opening.

Dilute 500X Track It™ Red DMSO stock solution (Component A) into Assay Buffer (Component B) to make a 10 to 25X Track It™ Red working solution. The working solution should be prepared enough for all the wells at 10 μL/well with the appropriate concentration. For example, to get a 10X final concentration of Track It™ Red for one 96-well microplate, dilute 20 μL of 500X Track It™ Red DMSO stock solution into 1 mL of Assay Buffer (Component B) to make 1 mL of 10X Track It™ Red working solution. Note: The final concentration of the Track It™ Red working solution should be empirically determined for different cell types and/or experimental conditions. It is recommended to test at the concentrations that are at least over a ten fold range.

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

1.  Add 10X Track It™ Red working solution to the cell wells which should be equal to 1/10 of the volume of cell culture medium. For example, for a 96-well plate, add 10 µL/well of 10X Track It™ Blue working solution into the cells.
   
   
2. Incubate the cells in a 37°C, 5% CO₂ incubator for 15 to 60 minutes.
   
   
3. Wash cells with Hanks and 20 mM Hepes buffer (HHBS) or an appropriate buffer.
   
   
4. Fill the cell wells with growth medium.
   
   
5. Analyze the cells using a fluorescence microscope with Texas Red filter or flow cytometer with 610/20 nm filter (PE-Texas Red channel).