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Synonyms: PD-MY 003, Hydroethidine
| Description | Dihydroethidium (PD-MY 003) is a peroxide indicator with blue fluorescence in the cytoplasm (λex=518 nm, λem=616 nm), which disappears when oxidized and is cell permeable. |
| In vitro | METHODS: Detection of cellular peroxides by Dihydroethidium: 1. Prepare a 5-10 mM stock solution of Dihydroethidium dye in DMSO, protect from light, and store in portions at -20℃. Dilute the stock solution with physiological buffers (e.g., PBS, HBSS, HEPES) to obtain a working solution of 5-20 µM Dihydroethidium dye. The optimal working concentration must be determined empirically. 3. Add the Dye Working Solution to the cells at a 1:1 ratio (100 µL of Dye Working Solution to 100 µL of Cell Suspension) and incubate for 5-60 minutes at room temperature or 37℃. 4. Wash with PBS and then perform fluorescence microscopy or flow cytometry. Dihydroethidium itself is blue fluorescent with a maximum excitation wavelength of 370 nm and a maximum emission wavelength of 420 nm, and after dehydrogenation, it binds to RNA or DNA to produce red fluorescence with a maximum excitation wavelength of 300 nm and a maximum emission wavelength of 610 nm, and 518 nm can be used as the excitation wavelength in practice. The above information is based on published literature. Experimental procedures should be appropriately modified to meet specific research demands. |
| Synonyms | PD-MY 003, Hydroethidine |
| Cell Research | Dihydroethidium detection of cell peroxides 1. Prepare 5-10 mM Dihydroethidium dye stock solution with DMSO, protect from light, and store in aliquots at -20°C. 2. Dilute the stock solution with physiological buffer (such as PBS, HBSS, HEPES) to obtain 5-20 µM Dihydroethidium dye working solution. The optimal working concentration must be determined empirically. 3. Add the dye working solution to the cells at a ratio of 1:1 (100 µL cell suspension to 100 µL dye working solution) and incubate at room temperature or 37°C for 5-60 minutes. 4. After washing with PBS, perform fluorescence microscopy or flow cytometry detection. Dihydroethidium itself is a blue fluorescent substance with a maximum excitation wavelength of 370 nm and a maximum emission wavelength of 420 nm. After dehydrogenation, it combines with RNA or DNA to produce red fluorescence with a maximum excitation wavelength of 300 nm and a maximum emission wavelength of 610 nm. In actual observation, 518 nm can also be used as the excitation wavelength. The above information is based on published literature. Experimental procedures should be appropriately modified to meet specific research demands. |
| Molecular Weight | 315.41 |
| Formula | C21H21N3 |
| Cas No. | 104821-25-2 |
| Smiles | CCN1C(c2ccccc2)c2cc(N)ccc2-c2ccc(N)cc12 |
| Relative Density. | 1.197g/cm3 |
| Storage | Keep away from direct sunlight Powder: -20°C for 3 years | In solvent: -80°C for 1 year Shipping with blue ice/Shipping at ambient temperature. | |||||||||||||||||||||||||||||||||||
| Solubility Information | DMSO: 125 mg/mL (396.31 mM), Sonication is recommended.  | |||||||||||||||||||||||||||||||||||
| In Vivo Formulation | 10% DMSO+40% PEG300+5% Tween 80+45% Saline: 0.5 mg/mL (1.59 mM), Solution. Please add the solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately. The formulation provided above is for reference purposes only. In vivo formulations may vary and should be modified based on specific experimental conditions. | |||||||||||||||||||||||||||||||||||
Solution Preparation Table | ||||||||||||||||||||||||||||||||||||
DMSO
Note : The dilution table applies only to solid products. For liquid products, please calculate the stock solution based on the stated concentration and/or density. | ||||||||||||||||||||||||||||||||||||
For example, if the intended dosage is 10 mg/kg for animals weighing 20 g , with a dosing volume of 100 μL per animal, and a total of 10 animals are to be administered, using a formulation of 
10% DMSO+ 40% PEG300+ 5% Tween 80+ 45% Saline/PBS/ddH2O , the resulting working solution concentration would be 2 mg/mL.Dissolve 2 mg of the compound in 100 μL DMSO to obtain a stock solution at a concentration of 20 mg/mL . If the required concentration exceeds the compound's known solubility, please contact us for technical support before proceeding.
1) Add 100 μL of the DMSO stock solution to 400 µL PEG300 and mix thoroughly until the solution becomes clear.
2) Add 50 µL Tween 80 and mix well until fully clarified.
3) Add 450 µL Saline,PBS or ddH2O and mix thoroughly until a homogeneous solution is obtained.
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