ROS assay cell type - human primary corneal epithelial cells

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

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Upstream tips
- Cells were pretreated with SiNPs at different concentrations (0, 25, 50, 100 μg/ml) for 24 h and 48 h.
Protocol tips
- Hydrogen peroxide 20 μM concentration was used for the assay.

- After adding DCFH, plate was incubated for 30min.
Protocol tips
- Cells were treated with 25 μM DCFDA in essential medium with 10% FBS in 37°C incubator for 45 min.

- Before analysing cells, they are treated with hyperosmotic media by adding NaCl for different time periods (30–180 min).
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