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Cat. No.: DHE-5 (for 5mg)

Description

Dihydroethidium, abbreviated as DHE, is the most commonly used fluorescent probe for detecting intracellular levels of superoxide anions. Its molecular formula is C21H21N3, and its molecular weight is 315.41.

Dihydroethidium is a widely used fluorescent probe for detecting superoxide anions. It can be directly used for labeling live cells. After being taken up by live cells, Dihydroethidium undergoes dehydrogenation in the presence of superoxide anions, resulting in the production of ethidium. Ethidium (e.g., ethidium bromide) can bind to RNA or DNA, generating red fluorescence. When the level of superoxide anions in the cell is high, more ethidium is produced, resulting in stronger red fluorescence. Conversely, when the level is low, the fluorescence is weaker. This allows for the detection of superoxide anion levels using Dihydroethidium.

Dihydroethidium itself emits blue fluorescence with a maximum excitation wavelength of 370nm and a maximum emission wavelength of 420nm. After dehydrogenation and binding to RNA or DNA, it emits red fluorescence with a maximum excitation wavelength of 300nm and a maximum emission wavelength of 610nm. When observing the fluorescence, an excitation wavelength of 535nm can also be used.

Dihydroethidium can be dissolved in DMSO to prepare an appropriate stock solution.

For intracellular detection of superoxide anions, the commonly used concentration of Dihydroethidium is 1-10μM. Typically, cells are incubated with an appropriate solution containing 0.5-5μM of Dihydroethidium at 37°C for approximately 30 minutes for probe loading. Subsequently, the cells can be washed, and fluorescence detection can be performed using flow cytometry or other suitable fluorescence detection instruments.

Additional Notes