Donor PBMCs were incubated with 5 µM CellTrace Blue Cell Proliferation kit (ThermoFisher, Cat # C34574) proliferation dye and then stimulated with anti-CD3/anti-CD28 beads (ThermoFisher, Cat # 11132D) in a 96-well round-bottom plate. Adenosine was added to appropriate wells day 0 at 1 mM final concentration and again on day 1 at 0.5 mM final concentration. Proliferation was assessed 4 days post-stimulation for proliferation dye dilution using flow cytometry.
Celltrace blue cell proliferation kit
Manufactured by Thermo Fisher Scientific
The CellTrace Blue Cell Proliferation kit is a fluorescent dye-based assay used to monitor cell division and proliferation in vitro. The kit provides a simple and reliable method for quantifying cell division by labeling cells with a blue-fluorescent dye that binds to cellular proteins. As cells divide, the dye is equally distributed between daughter cells, resulting in a progressive halving of the fluorescence intensity per cell with each division. This allows for the monitoring of cell proliferation over multiple generations.
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Lab products found in correlation
2 protocols using celltrace blue cell proliferation kit
1
Adenosine Modulates T Cell Proliferation
2
Cell Proliferation Assay with CellTrace Blue
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The CellTrace™ Blue cell proliferation kit (ThermoFisher, #C34568) was utilized to label 1 × 106 cells as per manufacturers protocol by incubation with 5 μM CellTrace™ Blue in protein- free medium for 20 min at 37 °C. Cells were washed with culture medium containing 1% FCS and seeded at a low density into replicate wells and maintained at 37 °C with 5% CO2. CellTrace™ Blue fluorescence (355ex /410em) and LC status GFP+/RFP+ was routinely assessed by microscopy using the Cytation™ 3 Multimode Imager (Biotek).
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