ROS‐Glo H2O2 Assay - Novel Luminescence-Based Assay for ROS Measurement

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.   This assay allows identification of conditions or test compounds, such as small molecule inhibitors or inducers, that alter ROS levels. The scalable multiwell format couples a stable luminescent signal to the level of H2O2 in a sample. An H2O2 Substrate is employed that reacts directly with H2O2 to generate a luciferin precursor. Upon addition of ROS-Glo™ Detection Reagent containing Ultra-Glo™ Recombinant Luciferase and D-Cysteine, the precursor is converted to luciferin by the D-Cysteine, and the produced luciferin reacts with Ultra-Glo™ Recombinant Luciferase to generate a luminescent signal that is proportional to H2O2 concentration. ROS-Glo™ H2O2 Assay Benefits:   Direct Cell-Based Detection - The assay can be performed in various cell culture media with or without serum, eliminating the need to remove media from cultured cells before performing the assay. Simple and Fast Protocol - The homogeneous assay is performed following a simple two-reagentaddition protocol that does not require sample manipulation. The assay can be completed in less than 2 hours. Automation Compatible - The assay is compatible with liquid handling robotics and can be scaled for use in multiwell formats. Non-HRP Based - 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. Flexible - 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.