Cellular ROS/Superoxide Detection Assay Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 1x10^4 cells were seeded for an assay.	- After 48h of culture, cells were treated with ROS/Superoxide Detection Mix for 30 min at 37°C

Upstream tips
- 1x10^4 cells were seeded for an assay.
Protocol tips
- After 48h of culture, cells were treated with ROS/Superoxide Detection Mix for 30 min at 37°C

Publication protocol

Free radicals and other reactive species were detected using the Cellular ROS/Superoxide Detection Assay Kit (Abcam) as recommended by the manufacturer. 1x104 RAW 264.7 macrophages per 96-well were seeded and treated as for the phagocytosis assay. After 48 hours, the culture medium was removed. The cells were then incubated with the ROS/Superoxide Detection Mix for 30 min at 37°C. After washing, Oxidative Stress (ROS; Green) and O2- (Red) -positive cells were counted and reported to the total amount of cell using the Zeiss Axiovert microscope. A minimum of 200 cells were counted per treatment condition and each treatment was tested in quadruplicate within a single experiment.

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