ROS-Glo™ H2O2 Assay

Description

The ROS-Glo™ H₂O₂ Assay is a homogeneous, fast and sensitive bioluminescent assay that measures the level of hydrogen peroxide (H₂O₂), a reactive oxygen species (ROS), directly in cell culture or in defined enzyme reactions. A derivatized luciferin substrate is incubated with sample and reacts directly with H₂O₂ to generate a luciferin precursor. Addition of ROS-Glo™ Detection Solution converts the precursor to luciferin and provides Ultra-Glo™ Recombinant Luciferase to produce light signal that is proportional to the level of H₂O₂ present in the sample.

Features – Benefits

• Direct Cell-Based Detection: The assay can be performed in various cell culture media with or without serum, eliminating the need to remove the media from cultured cells before performing the assay.
• Simple and Fast Assay: The homogeneous assay is performed following a simple two-reagent-addition protocol that does not require sample manipulation. The assay can be completed in less than 2 hours after reagent addition.
• Non-HRP-Based Detection: The ROS-Glo™ H₂O₂ Substrate reacts directly with H₂O₂, obviating the need for horseradish peroxidase (HRP) as a coupling enzyme and thus eliminating false hits associated with HRP inhibition.
• Automation-Compatible Format: Easily scale from 96- to 384-well plate formats.
• Flexible Assay: The assay can be used to screen compounds in both cell-based and enzyme-based formats.
• Multiplex-Compatible System: Get more informative data per well and reduce cell culture expenses by multiplexing with a real-time cytotoxicity assay (CellTox™ Green Cytotoxicity Assay) in the same well or with a viability assay.

Applications

• Measure changes in hydrogen peroxide levels directly in cell culture samples.
• Measure the activity of enzymes that generate or eliminate H₂O₂.
• Identify small molecule inhibitors or inducers that alter ROS levels either in cells in culture or in enzyme assays.

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