Promega Introduces Bioluminescent ROS-Glo H₂O₂ Assay for Enzymatic Screening Programs and Cell-based Assays

Promega Corporation announced today the launch of the ROS-Glo™ H_²O_² Assay. A non-horseradish peroxidase (HRP)-dependent, plate-based bioluminescent assay for detecting reactive oxygen species (ROS), ROS-Glo H_²O_² Assay is designed to specifically detect hydrogen peroxide (H_²O_²) in both enzyme and cell-based applications.

The ROS-Glo H_²O_² Assay measures the activity of enzymes that generate or eliminate H_²O_², and possesses many benefits over other commercially available assays for small molecule screening, including low false hit rate, signal stability, and compatibility with liquid handling. Importantly for screening applications, the assay does not rely on a reaction catalyzed by HRP, which is known to cause a high number of false hits. The assay can also be used to measure changes in the level of ROS by directly detecting H_²O_² in cultured mammalian cells after two reagent additions, with no cell sample preparation required, reducing variability and the number of cells required.

The new assays enable researchers to more effectively study the biology of ROS, and to screen large compound libraries for their capacity to alter H_²O_² levels in cultured cells or in enzymatic reactions. Although ROS can be beneficial to human cells as they are common byproducts of metabolism, they can also lead to cellular damage, such as oxidative stress, through environmental factors or aberrant metabolism.

The homogeneous ROS-Glo H_²O_² Assay uses a modified luciferin substrate that reacts directly with H₂O₂ to generate a luciferin precursor. Upon addition of detection reagent, precursor is converted to luciferin and Ultra-Glo™ Recombinant Luciferase included in the detection reagent produces a light signal proportional to the level of H_²O_² in the sample.

Due to the bioluminescent format and high sensitivity, ROS-Glo H_²O_² Assay can be multiplexed with a variety of other cell-based assays to gain additional data from a sample. For example, CellTox™ Green Cytotoxicity Assay reagent can be added to the same well at cell plating or treatment to kinetically monitor cytotoxicity by measuring changes in fluorescence, followed by implementation of the ROS-Glo H_²O_² Assay to measure changes in ROS levels by luminescence detection.