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.
P450 Induction in Cryopreserved Hepatocytes from PXR and CAR Nuclear Receptor Knock-out RatsPoster
The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are closely related transcription factors that regulate the expression of phase I (cytochrome P450s), phase II metabolizing enzymes and transporter genes in response to xenobiotics, including prescription drugs.READ MORE
Knockout of microRNAs Using the CRISPR-Cas9 System with Paired Synthetic crRNAsPoster
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE
A New Method for Analyzing MSe/All Ions Fragmentation in Xenobiotic Metabolism StudiesPoster
During early drug discovery, the study of metabolism plays an essential role in determining which drug candidates move forward into development and later stages. As an alternative to traditional Data Dependent Acquisition (DDA), the use of MSE/All Ions Fragmentation (AIF) has become common in metabolite identification workflows for the analysis of metabolic hot spots. Here we present a solution for analysis of MSE/AlF in metID studies.READ MORE