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Detection of ROS in Jurkat cells. Jurkat cells were seeded on the same day at 300000 cells/100 uL/well in a Costar black wall/clear bottom 96-well plate. The ROS Red assay stain solution (100 uL/well) was added and incubated in a 5% CO2, 37°C incubator for 1 hour. The cells were treated with or without 1mM H2O2 for 2 hours. The fluorescence signal was monitored at Ex/Em = 520/605 nm (cut off = 590 nm) with bottom read mode using FlexStation.