ROS were measured as previously described (22 (link)). BALF cells were washed with PBS. To measure intracellular ROS, cells were incubated for 10 min at room temperature with PBS containing 3.3 µM 2′,7′-dichlorofluorescein (DCF) diacetate (Molecular Probes, Eugene, OR, USA), to label intracellular ROS. DCF-stained cells (1×104) were subjected to fluorescence-activated cell sorting analysis to measure ROS levels using a FACSCalibur instrument (BD Biosciences, San Jose, CA, USA). The data were analyzed with CellQuest Pro 3.3 software (BD Biosciences).
2 7 dichlorofluorescein dcf diacetate
Manufactured by Thermo Fisher Scientific
Sourced in United States
2',7'-dichlorofluorescein (DCF) diacetate is a fluorogenic probe used for detecting the presence of reactive oxygen species (ROS) in cells. It is a nonfluorescent compound that can passively diffuse into cells, where it is hydrolyzed by intracellular esterases to the fluorescent compound 2',7'-dichlorofluorescein (DCF), which can then be detected using a fluorescence-based assay.
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2 protocols using 2 7 dichlorofluorescein dcf diacetate
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Intracellular ROS Measurement in BALF Cells
2
Intracellular ROS Measurement in BAL Cells
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ROS was measured using a previously described method [21 (link)]. BAL cells were washed with PBS. To measure intracellular ROS, cells were incubated for 10 min at room temperature with PBS containing 3.3 μM 2′,7′-dichlorofluorescein (DCF) diacetate (Molecular Probes, Eugene, OR, USA) to label intracellular ROS. We performed fluorescence-activated cell sorting analysis with DCF stained cells (1 × 104 cells) in BALF to measure ROS levels using a FACSCalibur (BD Biosciences, San Jose, CA, USA). The data were analysed using CellQuest Pro (BD Biosciences).
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