Reactive Oxygen Species Assay Kit for Superoxide Anion (DHE)

The Target Mol Superoxide Anion Reactive Oxygen Species (DHE) detection kit is a fluorescent probe using DHE to detect intracellular superoxide anion levels. Superoxide anions (· O2-) are negatively charged reactive oxygen species formed by oxygen molecules acquiring an electron. They are strong oxidants and can react with various biomolecules. In living organisms, it is mainly produced by the metabolic processes of mitochondrial respiratory chain and enzymatic reactions mediated by NADPH oxidase, and can also be induced by environmental stimuli such as UV and drug metabolism. When the level of superoxide anions is moderate, it participates in normal physiological activities such as immune defense and cell signaling. Excessive levels can trigger oxidative stress, causing cell damage and being associated with the occurrence of various diseases.
Dihydroethium (DHE) is a lipid soluble fluorescent probe that can freely penetrate the cell membrane and enter the cell. The principle of DHE detection of superoxide anions is that when DHE does not bind to superoxide anions, it emits weak blue fluorescence (maximum excitation wavelength is 370nm, maximum emission wavelength is 420nm). When combined with intracellular superoxide anions, DHE undergoes specific oxidation reactions and is oxidized to produce Ethidium with strong fluorescent properties. Ethidium can bind to DNA or RNA and emit red fluorescence upon excitation (maximum excitation wavelength is 300nm, maximum emission wavelength is 610nm, and 535nm can be used as the excitation wavelength for actual observation). The intensity of red fluorescence is positively correlated with the concentration of superoxide anions, and the level of superoxide anions in cells can be analyzed by detecting the fluorescence intensity.
This kit is suitable for various detection instruments such as fluorescence microscope, laser confocal microscope, fluorescence spectrophotometer, and flow cytometer. If this reagent kit is used for a 6-well plate with a detection volume of 1 mL per well, it can detect 100 times; If used for a 96 well plate with a detection volume of 100 µ L per well, it can be tested 1000 times.