Description
• Measures total reactive oxygen species and reactive nitrogen species, including hydrogen peroxide, nitric oxide, peroxyl radical, and peroxynitrite anion
• Suitable for use with serum, plasma, urine, cell lysates or cell culture supernatants
• Detection sensitivity limit of 10 pM for DCF and 40 nM for hydrogen peroxide
The OxiSelect In Vitro ROS/RNS Assay provides a sensitive method to detect total reactive oxygen species (ROS) plus reactive nitrogen species (RNS) in a wide variety of sample types. This assay employs a proprietary fluorogenic probe, DCFH-DiOxyQ; the probe is primed with a dequenching reagent to the highly reactive DCFH form. In the presence of ROS and RNS, the DCFH is rapidly oxidized to the highly fluorescent DCF.
Introduction: Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are well-established molecules responsible for the deleterious effects of oxidative stress. Accumulation of free radicals coupled with an increase in oxidative stress has been implicated in the pathogenesis of several disease states. The role of oxidative stress in vascular diseases, diabetes, renal ischemia, atherosclerosis, pulmonary pathological states, inflammatory diseases, cancer, as well as ageing has been well established. Free radicals and other reactive species are constantly generated in vivo and cause oxidative damage to biomolecules, a process held in check by the existence of multiple antioxidant and repair systems as well as the replacement of damaged nucleic acids, proteins and lipids. Measuring the effect of antioxidant therapies and ROS/RNS activity is crucial to suppressing or treating oxidative stress inducers.
Introduction
The OxiSelect In Vitro ROS/RNS Assay Kit is an assay for measuring the total free radical presence of a sample. The assay employs a proprietary quenched fluorogenic probe, dichlorodihydrofluorescin DiOxyQ (DCFH-DiOxyQ), which is a specific ROS/RNS probe that is based on similar chemistry to the popular 2', 7'-dichlorodihydrofluorescein diacetate. The DCFH-DiOxyQ probe is first primed with a quench removal reagent, and subsequently stabilized in the highly reactive DCFH form. In this reactive state, ROS and RNS species can react with DCFH, which is rapidly oxidized to the highly fluorescent 2', 7'-dichlorodihydrofluorescein (DCF) (Figure 1). Fluorescence intensity is proportional to the total ROS/RNS levels within the sample. The DCFH-DiOxyQ probe can react with hydrogen peroxide (H2O2), peroxyl radical (ROO?), nitric oxide (NO), and peroxynitrite anion (ONOO-). These free radical molecules are representative of both ROS and RNS, thus allowing for measurement of the total free radical population within a sample. OxiSelect In Vitro ROS/RNS Assay Kit can also be used to evaluate antioxidant's effect on free radicals. The kit has a detection sensitivity limit of 10 pM for DCF and 40 nM for H2O2 respectively. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples