DCF ROS/RNS Assay Kit (biofluids, culture supernatant, cell lysates)

Product details

How the assay works

DCF ROS/RNS Assay Kit (biofluids, culture supernatant, cell lysates)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.

DCF ROS/RNS Assay Kit (biofluids, culture supernatant, cell lysates)protocol summary:

- Unknown ROS or RNS samples or standards are added to the wells with a catalyst that helps accelerate the oxidative reaction.
- After a brief incubation, the prepared DCFH probe is added to all wells and the oxidation reaction is allowed to proceed
- Samples are measured fluorometrically against a hydrogen peroxide or DCF standard.

How other researchers are using

DCF ROS/RNS Assay Kit (biofluids, culture supernatant, cell lysates) has been used in variety of sample type including:
- Monkey kidney epithelial cells 1
- Rat cardiomyocyte-derived H9c2 cells 2
- Human MSCs and human fibroblasts 3

References:
1 - Velбsquez-Torres M et al. 2023; 2 - Zhao H et at 2023; 3 - Choudhury D et al. 2022.

Related and recommended products

See alternative ROS assays available in orange (ab186028), red (ab186027), and deep red (ab186029).

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.