Life Technologies
CellTrace™ Far Red Cell Proliferation Kit, for flow cytometry
C34564
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This cell labeling kit was chosen as an alternative to CFSE and can be detected in the APC channel. The same protocol used with CFSE was applied and I observed comparable results between CFSE and this cell proliferation kit.
Cell proliferation
Purified CD4 T cells
1. Purified CD4 T cells isolated using magnetic beads were washed with PBS and resuspended to 20 million cells/ml in PBS. 2. Celltrace Far Red was reconstituted in 18ul of DMSO to make a 5mM stock. 3. For in vivo cell proliferation, Celltrace Far Red was diluted to 10uM stock in PBS. 4. Equal volume of diluted Celltrace Far Red was added to the cells to give a final concentration of 5uM. 5. Cells were incubated at a 37C waterbath for 30 mins. 6. Neat fetal calf serum was added to the cells to quench Celltrace Far Red and stop the reaction. 7. Cells were washed with PBS and now ready for adoptive transfer into mice.
Do not exceed the 30min incubation at 37C waterbath. To detect Celltrace Far Red positive cells prior to adoptive transfer, you might have to lower the voltage on the cytometer in the APC channel to pick positive cells.
Very bright staining of positive cells. Good alternative to CFSE if you want to free up the FITC/Alexa488 channel on the cytometer. Minimal overlap with CFSE and can be used together with CFSE-labeled cells to create a mix population for adoptive transfers.
None
Bright staining
Voltage adjustments on the cytometer might be needed
Highly recommended.
Product Name: CellTrace Far Red Cell Proliferation KitSupplier Page