| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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Dihydroethidium (formerly known as Hydroethidine and PD-MY 003) is a cell-permeable blue fluorescent dye and a superoxide indicator which exhibits blue-fluorescence in the cytosol until oxidized, where it intercalates within the cell's DNA, staining its nucleus with a bright fluorescent red. DHE-derived fluorescence at 570 nm is a convenient method to detect intracellular and extracellular superoxide produced by phagocytic and vascular NADPH oxidase. In addition, Dihydroethidium is also reported to be neuroprotective by reducing superoxide in mice after stroke.
| ln Vitro |
General procedure: 1.1 Prepare the stock solution by combining 1 mg of dihydroethidium with 0.31 mL of DMSO to create 10 mM of the compound. Note: To prevent repeated freezing and thawing, it is advised that the stock solution be stored at -20°C or -80°C in the dark. 1.2 Dihydroethidium working solution preparation: Dilute 1–10 μM of the working solution in PBS or serum-free cell culture medium. Note: Please modify the Dihydroethidium working fluid concentration based on the current circumstances. Section 2.1: Cell staining 2.1: Centrifuge 1000g of suspended cells for 3–5 minutes at 4°C; discard supernatant. Use PBS to wash twice, for five minutes each time. Adherent cells: To isolate the cells and create a single cell suspension, discard the cell culture medium and add trypsin. Discard the supernatant after centrifuging at 1000g for three to five minutes at 4°C. Use PBS to wash twice, for five minutes each time. 2.2 Add 1 milliliter of the Dihydroethidium working solution, then let it sit at room temperature for half an hour. 2.3 Discard the supernatant after centrifuging at 400 g for three to four minutes at 4°C. 2.4 Use PBS twice, giving each wash five minutes. 2.5 Re-suspend the cells in PBS or serum-free cell culture medium, then use a flow cytometer or fluorescence microscope to observe them. Storage for a year at -20°C with protection from light Precautions 1. The original solution should be kept refrigerated at -20°C or -80°C, shielded from light, and not frozen and thawed too frequently. 2. Please modify the Dihydroethidium working fluid concentration based on the current circumstances. 3. This item is intended for use in research and development. 4. When working, please wear a lab coat and disposable gloves for your own health and safety.
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| ln Vivo |
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| Animal Protocol |
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| References |
[1]. Zielonka J, et al. Global profiling of reactive oxygen and nitrogen species in biological systems: high-throughput real-time analyses. J Biol Chem. 2012 Jan 27;287(5):2984-95.
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| Molecular Formula |
C21H21N3
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| Molecular Weight |
315.41
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| CAS # |
104821-25-2
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| Related CAS # |
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| SMILES |
NC1=CC=C2C3=C(C=C(N)C=C3)C(C4=CC=CC=C4)N(CC)C2=C1
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| Synonyms |
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.93 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1705 mL | 15.8524 mL | 31.7048 mL | |
| 5 mM | 0.6341 mL | 3.1705 mL | 6.3410 mL | |
| 10 mM | 0.3170 mL | 1.5852 mL | 3.1705 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
 Oxidation of HE by O2˙̄,•NO, and co-generated•NO and O2˙̄and global profiling of products.J Biol Chem.2012 Jan 27;287(5):2984-95. |
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 Real-time monitoring of products of HE oxidation by activated macrophages.J Biol Chem.2012 Jan 27;287(5):2984-95. |
 Global profiling of H2O2and ONOO−-derived oxidants by monitoring the oxidation of Amplex® Red.J Biol Chem.2012 Jan 27;287(5):2984-95. |
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