Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader
Product nameDCF ROS/RNS Assay Kit (biofluids, culture supernatant, cell lysates)
DCF ROS/RNS Assay Kit (biofluids, culture supernatant, cell lysates) (ab238535) 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). 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-).
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.
Review the oxidative stress marker and assay guide, or the full metabolism assay guide to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.
To measure reactive oxygen species within cells, we recommend DCFDA / H2DCFDA - Cellular ROS Assay Kit ab113851. Alternative ROS assays are available in orange (ab186028), red (ab186027), and deep red (ab186029). ab238535 is used to measure ROS in biofluids, culture supernatants and cell lysates.
For assays designed to differentiate ROS, superoxides, and reactive nitrogen species: to assay ROS and superoxides use ab139476; to assay ROS, superoxides, and reactive nitrogen species use ab139473; to assay superoxides use ab219943.
Storage instructionsPlease refer to protocols.
Components 96 tests Catalyst (250X) 1 x 20µl DCF Standard 1 x 100µl DCF-DiOxyQ 1 x 50µl Hydrogen Peroxide 1 x 100µl Priming Reagent 1 x 250µl Stabilization Solution (10X) 1 x 1.5ml
ab238535 has been referenced in 2 publications.
- Vandierendonck A et al. NOX1 inhibition attenuates the development of a pro-tumorigenic environment in experimental hepatocellular carcinoma. J Exp Clin Cancer Res 40:40 (2021). PubMed: 33485364
- Cao JY et al. Autophagosome protects proximal tubular cells from aldosterone-induced senescence through improving oxidative stress. Ren Fail 43:556-565 (2021). PubMed: 33757397