The effect of Cu-CDs treatments on the redox homeostasis of breast cancer cells was further analyzed by assessing ROS production in MDA-MB-231 cells.48 (link) MDA-MB-231 cells were incubated with 10 μM dichlorofluorescein diacetate (DCFH-DA) staining solution (S0033, Beyotime, Shanghai, China) at 37 °C for 30 min. After DAPI staining, the DCFH-DA staining was examined and recorded with the ROS production level analyzed by the quantification of DCFH-DA staining intensity by Image J software.
Dcfh da staining solution
Manufactured by Beyotime
Sourced in China
DCFH-DA (2',7'-Dichlorofluorescin diacetate) is a cell-permeant fluorogenic probe commonly used for the detection and quantification of intracellular reactive oxygen species (ROS). Upon cellular uptake, DCFH-DA is cleaved by intracellular esterases and then oxidized by ROS to produce the fluorescent dye 2',7'-dichlorofluorescein (DCF), which can be detected and measured using fluorescence techniques.
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2 protocols using dcfh da staining solution
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Assessing ROS in Breast Cancer Cells
2
Measuring Cellular ROS Production by DCFH-DA Assay
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ROS production in cells was determined by dichloro-dihydro-fluorescein diacetate (DCFH-DA). Cells were incubated with 500 µl of a DCFH-DA-staining solution (Beyotime Biotechnology, China) at 37 °C for 20 min. Then, the staining solution was discarded and cells washed with 3 changes of serum-free medium. Accuri C6 flow cytometry (BD, biosciences, USA) was performed to detect the fluorescence intensity of each group. The excitation wavelength was 488 nm, and the emission wavelength was 525 nm. The experiment was repeated 3 times for each group.
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