Quantifying Cellular Reactive Oxygen Levels

The DCFDA Cellular Reactive Oxygen Species Detection Assay Kit (ab113851; Abcam) was used to estimate the reactive oxygen species (ROS) levels in cells [57 (link)]. Briefly, 2.5 x 10⁴ cells per well were seeded in dark, clear bottom 96-well microplates and allowed to adhere overnight. Then, the cells were incubated with 25 μM DCFDA for 1 h at 37°C. The fluorescence was analyzed using a fluorescent plate reader at an excitation wavelength at 485 nm and an emission wavelength at 535 nm [58 (link)].

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Wang Y., Yang Q., Shen S., Zhang L., Xiang Y, & Weng X. (2020). Mst1 promotes mitochondrial dysfunction and apoptosis in oxidative stress-induced rheumatoid arthritis synoviocytes. Aging (Albany NY), 12(16), 16211-16223.

Publication 2020

DCFDA Cellular Reactive Oxygen Species Detection Assay Kit

Assay Cells Dcfda Fluorescence Reactive oxygen species

Corresponding Organization :

Other organizations : Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Capital Medical University, Beijing Tongren Hospital

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Variable analysis

independent variables

DCFDA concentration (25 μM)

dependent variables

Reactive oxygen species (ROS) levels

control variables

Cell seeding density (2.5 x 10^4 cells per well)
Incubation time (1 h)
Incubation temperature (37°C)
Microplate type (dark, clear bottom 96-well)

controls

No information provided about positive or negative controls.

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