Dihydroethidium

Catalog No.S6792 Batch:S679201

Technical Data

Formula	C₂₁H₂₁N₃
Molecular Weight	315.41	CAS No.	104821-25-2
Solubility (25°C)*	In vitro	DMSO	63 mg/mL (199.74 mM)
		Ethanol	2 mg/mL (6.34 mM)
		Water	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description	Dihydroethidium (DHE, HE, Hydroethidine, PD-MY 003) is a cell-permeable blue fluorogenic probe used for detecting intracellular superoxide radical anion.
In vitro	1.1 Preparation of the stock solution Dissolve 1 mg of Dihydroethidium in 0.31 mL of DMSO to obtain 10 mM of Dihydroethidium. Note: It is recommended to store the stock solution at -20°C or -80°C away from light and avoid repetitive freeze-thaw cycles. 1.2 Preparation of Dihydroethidium working solution Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of Dihydroethidium working solution. Note: Please adjust the concentration of Dihydroethidium working solution according to the actual situation. Cell staining 2.1 For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time. For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time. 2.2 Add 1 mL of Dihydroethidium working solution, and then incubate at room temperature for 30 minutes. 2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant. 2.4 Wash twice with PBS, 5 minutes each time. 2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

Description

Dihydroethidium (DHE, HE, Hydroethidine, PD-MY 003) is a cell-permeable blue fluorogenic probe used for detecting intracellular superoxide radical anion.

In vitro

1.1 Preparation of the stock solution
Dissolve 1 mg of Dihydroethidium in 0.31 mL of DMSO to obtain 10 mM of Dihydroethidium.
Note: It is recommended to store the stock solution at -20°C or -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Dihydroethidium working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of Dihydroethidium working solution.
Note: Please adjust the concentration of Dihydroethidium working solution according to the actual situation.
Cell staining
2.1 For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of Dihydroethidium working solution, and then incubate at room temperature for 30 minutes.
2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

Protocol (from reference)

References

[1] Jacek Zielonka, B Kalyanaraman. Free Radic Biol Med. 2010 Apr 15;48(8):983-1001.

Selleck's Dihydroethidium has been cited by 3 publications

Dual-imaging magnetic nanocatalysis based on Fenton-like reaction for tumor therapy [ J Mater Chem B, 2022, 10(18):3462-3473]	PubMed: 35403639
Mechanism for enhanced transduction of hematopoietic cells by recombinant adeno-associated virus serotype 6 vectors [ FASEB J, 2020, 34(9):12379-12391]	PubMed: 32960474
EZH2 Inhibition Ameliorates Transverse Aortic Constriction-Induced Pulmonary Arterial Hypertension in Mice [ Can Respir J, 2018, 2018:9174926]	PubMed: 29854032

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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