ROS-Glo™ H2O2 Assay: A Luminescent Assay for Detection of Reactive Oxygen Species Poster
Poster Oct 09, 2015
Gediminas Vidugiris, Sarah Duellman, John Shultz, Jolanta Vidugiriene, Hui Wang, Jean Osterman, Wenhui Zhou, Poncho Meisenheimer and James J. Cali
H2O2 is a reactive oxygen species (ROS) that is measured in cells as a marker of oxidative stress. It is also measured as a marker of enzyme activities that either consume or produce H2O2. It is desirable to screen chemical compounds for their capacity to alter H2O2 levels in cultured cells or for their effects on H2O2 levels in enzyme reactions. Current fluorescent assay formats are prone to false hit rates that are too high for efficient screening applications. The ROS-Glo™ luminescent H2O2 assay detects H2O2 directly, minimizes false hit rate and provides simple formats for cell-based and enzymatic assays.
Since various ROS are interconverted to H2O2 in the cell and H2O2 is the longest lived ROS, an increase in H2O2 can reflect a general increase in the ROS level. Our method for measuring hydrogen peroxide utilizes the H2O2 Substrate, which directly reacts with H2O2 to produce a luciferin precursor. Addition of the ROS-Glo™ Detection Solution converts the precursor to luciferin and provides luciferase and other components to produce a light signal proportional to the level of H2O2.
Primary Human Hepatocyte (PHH) culture provides the closest in vitro model to human liver that can produce a metabolic profile of a given drug very similar to that found in vivo. Recently we have developed an easy-to-assemble user-friendly in vitro Primary Human Hepatocyte (PHH) 3D-spheroid modelREAD MORE