OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells were pretreated with SiNPs at different concentrations (0, 25, 50, 100 μg/ml) for 24 h and 48 h.	- Hydrogen peroxide 20 μM concentration was used for the assay. - After adding DCFH, plate was incubated for 30min.

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.

Publication protocol

Generation of ROS was detected using OxiSelect In Vitro ROS/RNS Assay Kit (catalog number: STA-347: Cell Biolabs. Inc., San Diego, CA, USA). HCEC were treated with each size of SiNPs at different concentrations (0, 25, 50, 100 μg/ml) for 24 h and 48 h. Following incubation, supernatants and cells were collected separately. Sonicated cells and the supernatants were assayed for the measurement of intracellular and extracellular ROS respectively following the manufacturer’s protocol. Hydrogen peroxide (20 μM) was used to generate a standard curve by serial dilution and 50 μL of appropriate samples from cells or supernatants were transferred to a 96-black well plate. 50 μL of Catalyst and 100 μL of dichloro-dihydro-fluorescein diacetate (DCFH) solution were added in order and the plate was incubated at room temperature for 30 min. Finally, the fluorescence was measured at 480 nm excitation/530 nm emission.

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Manufacturer protocol

Download the product protocol from Cell Biolabs for OxiSelect™ In Vitro ROS/RNS Assay Kit (Green Fluorescence) below.

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