(1365-B) Reactive oxygen species detection using a luminescent cell-based assay
Monday, February 5, 2024
2:00 PM – 3:00 PM EST
Location: Exhibit Halls AB
Abstract: Reactive oxygen species (ROS) are chemically reactive derivatives of molecular oxygen that arise from aerobic metabolism. ROS play an important role in cell signaling and gene expression. However, if ROS accumulate at high levels inside the cell, they can also damage DNA, RNA, proteins and lipids. High levels of ROS can occur due to oxidative stress and pose an overall threat to cellular health. ROS are also associated with aging, apoptosis or necrosis, and are implicated in diabetes, vascular diseases, pulmonary disorders, inflammatory diseases, and cancer.
There are many intracellular sources of ROS, including mitochondria and NADPH oxidases. ROS detection assays are used in drug screening campaigns to determine the effects of compounds from chemical libraries on enzymatic reactions (e.g. NADPH oxidase) and to study the effects of antioxidant therapies. ROS typically have a short half-life. H2O2 is an exception and has the longest half-life of all known ROS, which makes it a good marker of oxidative stress and well suited for ROS detection.
Here we show the results of cellular production of hyrdogen peroxide detect with Promegas ROS-Glo assay using both positive and negative regulators of ROS generation. Menadione is a compound that interrupts the electron transport chain in mitochondria, producing large amounts of ROS in cultured cells and is used as a positive control reagent in ROS detection. Conversely, N-Acetyl-L-cysteine (NAC) functions as an antioxidant and shows an inhibitory effect on ROS generation. The assay was detected with high sensitivity thus enabling miniaturization.