DCFDA - Cellular Reactive Oxygen Species Detection Assay Kit

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	- After adding DCFDA 10 μM, cells were incubated for 30 min at 37°C in the dark

Protocol tips
- After adding DCFDA 10 μM, cells were incubated for 30 min at 37°C in the dark

Publication protocol

Intracellular ROS levels were quantified by using the DCFDA (2′,7′-dichloro fluorescein diacetate)-Cellular Reactive Oxygen Species Detection Assay Kit (Abcam, UK). DCFDA was oxidized by ROS in viable cells to 2′,7′-dichloro fluorescein (DCF), which is highly fluorescent at 530 nm. The cells were washed three times with phosphate-buffered saline (PBS). DCFDA, diluted to a final concentration of 10 μM, was added, and the cells were incubated for 30 min at 37°C in the dark. After washing three times with PBS, fluorescence was measured with a multimode microplate reader (Tecan Trading AG, Switzerland) at excitation and emission wavelengths of 488 and 525 nm, respectively. ROS level was calculated as the absorbance ratio of experimental and control cells and expressed as a percentage.

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