ROS-Glo™ H2O2 Assay

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells were plated at a density of 100,000 cells/well	- plates were placed in hyperoxia or normoxia for 0 h, 4 h, 8 h, 12 h and 24 h. H2O2 substrate solution was added to the plates 6 h. - The detection solution was added to the plates 20 min before each read.

Upstream tips
- Cells were plated at a density of 100,000 cells/well
Protocol tips
- plates were placed in hyperoxia or normoxia for 0 h, 4 h, 8 h, 12 h and 24 h. H2O2 substrate solution was added to the plates 6 h. - The detection solution was added to the plates 20 min before each read.

Publication protocol

The ROS-Glo™ H2O2 Assay (Promega Inc.) was used to measure the level of hydrogen peroxide (H2O2), a reactive oxygen species (ROS), directly in cell culture according to the manufacturer's recommendations. Cells were plated at a density of 100,000 cells/well on a 96 well format plate, and incubated overnight for attachment before transfection with respective siRNA. The plates were placed in hyperoxia or normoxia for 0 h, 4 h, 8 h, 12 h and 24 h. H2O2 substrate solution was added to the plates 6 h before read, and the plates were replaced in hyperoxia chamber. The detection solution was added to the plates 20 min before each read and incubated at room temperature and relative luminescence was recorded using SpectraMaxM3 microplate reader (Molecular Devices LLC, Sunnyvale, CA).

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Manufacturer protocol

Download the product protocol from Promega for ROS-Glo™ H2O2 Assay below.

Download manufacturer protocol

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