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Intracellular calcium response of Jurkat cells to stimulation with A23187. Jurkat cells were stained with 5 μM BD Pharmingen™ Fluo-4 AM (Cat. No. 565878) in DPBS with calcium and magnesium for 20 minutes at 37°C. Cells were pelleted once and further incubated in complete medium for 30 minutes to allow complete hydrolysis of AM moieties. Cells were then pelleted and resuspended in DPBS with calcium and magnesium and analyzed on an BD Accuri™ C6 in the FL-1 channel. After 30 seconds of acquisition, 1 μM calcium ionophore A23187 (left, Sigma Aldrich Cat. No. C7522) or DMSO vehicle (right) was added to the tube, followed by mixing with the pipette. Jurkat cells show an increase in intracellular calcium concentration with addition of calcium ionophore A23187, but not with addition of DMSO vehicle.
BD Pharmingen™ Fluo-4 AM
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