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

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Before treating cells with DCFH-DA treatment for 60 min at 37°C cells were washed with prechilled PBS.

Protocol tips
- Before treating cells with DCFH-DA treatment for 60 min at 37°C cells were washed with prechilled PBS.

Publication protocol

To examine whether the production of ROS induced by ox-LDL treatment could be blocked by QCT in macrophages, RAW264.7 cells were subjected to the treatment of QCT (20 μM) for 24 h followed by ox-LDL (50 μg/ml) treatment for another 24 h. ROS production was detected as described previously (Wang et al., 2015). As non-fluorescent DCFH-DA could be de-esterified intracellularly and turns into fluorescent 2′,7′-dichlorofluorescin (DCFH) upon oxidation by free radicals, cells pellets were washed with prechilled PBS and processed for DCFH-DA treatment for 60 min at 37°C. Subsequently, ROS production in response to oxidation in macrophages was detected by spectrofluorimetry, which exciting light at 488 nm and emitting light at 525 nm.

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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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