Reactive oxygen species (ROS) is a by-product of biological aerobic metabolism, including oxygen ions, peroxides, and oxygen-containing free radicals. More and more studies have proved that ROS is a messenger molecule of multiple signal pathways, triggered or directly related to cancer, neurodegenerative diseases, fibrosis, cardiovascular and other diseases. Monitoring the level of ROS will help clarify the mechanism of mitochondrial disease and introduce new treatments.
Figure 1. Schematic localization of the cellular production of ROS and RNS. (Cottet R. C., et al., 2011)
Creative Biogene provides customers around the world with a variety of ROS determination methods, including ESR determination, DCFH-DA fluorescent probe detection, NBT staining, DAB staining, and quantitative determination of hydrogen peroxide. Here we only introduce ESR determination and DCFH-DA fluorescent probe detection scheme. Please contact us for other techniques that interest you.
Dichlorofluoresceindiacetate (DCFH-DA) is a ROS probe, which undergoes deacetylation in cells, followed by ROS-mediated oxidation to produce fluorescent substances (excitation wavelength λex=485nm, emission wavelength λem=530nm). DCFH-DA can be used to measure the production of ROS in cytoplasm and organelles (such as mitochondria). The fluorescence intensity can be measured quantitatively by a microplate spectrophotometer.
Electron spin resonance (ESR) is a spectroscopy that observes electron spin resonance by irradiating samples with microwaves. ESR spectroscopy can be used to analyze the existence and quantification of unpaired electrons, as well as the atomic states around unpaired electrons. Besides, its sensitivity is in principle 700times higher than that of NMR which uses a magnetic field to detect nuclear spin. By using DMPO to supplement hydroxyl radical (HO∙), antioxidants can be evaluated from the spectrum before and after the injection of antioxidants.
Creative Biogene always provides you with customized services for mitochondrial research, and our one-stop research and analysis platform can meet all your research needs.
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