Hydroethidine [Dihydroethidium] *CAS 104821-25-2*

Hydroethidine

Dihydroethidium; CAS 104821-25-2

Hydroethidine operates effectively as a probe for measurement of reactive oxygen species. The dye enters cells freely and is dehydrogenated to an ethidium compound. The probe has been used extensively with NK cell and as a vital dye for identification of proliferation and hypoxic cells in tumors. Studies have been performed using neutrophils and endothelial cells as well as HL60 cells and macrophages. A major advantage of this probe is its ability to distinguish between superoxide and H2O2. Fluorescence emission occurs at around 600 nm.

Protocol

COA

Catalog	Size	Price	Quantity
15200	25 mg	Price
15202	5X1 mg	Price

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Hydroethidine [Dihydroethidium] *CAS 104821-25-2* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	317.048 µL	1.585 mL	3.17 mL	15.852 mL	31.705 mL
5 mM	63.41 µL	317.048 µL	634.095 µL	3.17 mL	6.341 mL
10 mM	31.705 µL	158.524 µL	317.048 µL	1.585 mL	3.17 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	315.41 g/mol	=	mL	x	mM	=	mmol

Citations

View all 17 citations: Citation Explorer

Authors:

Nikolova, Biliana and Semkova, Severina and Tsoneva, Iana and Stoyanova, Elena and Lefterov, Pavel and Lazarova, Dessislava and Zhelev, Zhivko and Aoki, Ichio and Higashi, Tatsuya and Bakalova, Rumiana

Journal:

Anticancer Research (2020): 5159--5170

Bifunctional nanozyme activities of layered double hydroxide derived Co-Al-Ce mixed metal oxides for antibacterial application

Authors:

Chen, Chao and Wang, Yi and Zhang, Dun

Journal:

Journal of Oceanology and Limnology (2020): 1--13

Dual response mimetic enzyme of novel Co4S3/Co3O4 composite nanotube for antibacterial application

Authors:

Wang, Jin and Wang, Yi and Zhang, Dun and Xu, Chaojie and Xing, Ronge

Journal:

Journal of Hazardous Materials (2020): 122278

Exploring the bactericidal performance and application of novel mimic enzyme Co4S3

Authors:

Wang, Jin and Wang, Yi and Zhang, Dun

Journal:

Journal of colloid and interface science (2019)

Bifunctionalized Novel Co-V MMO Nanowires: Intrinsic Oxidase and Peroxidase Like Catalytic Activities for Antibacterial Application

Authors:

Wang, Yi and Chen, Chao and Zhang, Dun and Wang, Jin

Journal:

Applied Catalysis B: Environmental (2019): 118256

References

View all 24 references: Citation Explorer

The confounding effects of light, sonication, and Mn(III)TBAP on quantitation of superoxide using hydroethidine

Authors:

Zielonka J, Vasquez-Vivar J, Kalyanaraman B.

Journal:

Free Radic Biol Med (2006): 1050

Pulse radiolysis and steady-state analyses of the reaction between hydroethidine and superoxide and other oxidants

Authors:

Zielonka J, Sarna T, Roberts JE, Wishart JF, Kalyanaraman B.

Journal:

Arch Biochem Biophys. (2006)

Generation of superoxide anion by equine spermatozoa as detected by dihydroethidium

Authors:

Burnaugh L, Sabeur K, Ball BA.

Journal:

Theriogenology. (2006)

Analysis of dihydroethidium-derived oxidation products by HPLC in the assessment of superoxide production and NADPH oxidase activity in vascular systems

Authors:

Fern, undefined and es DC, Wosniak J, Pescatore LA, Bertoline MA, Liberman M, Laurindo F, Santos CX.

Journal:

Am J Physiol Cell Physiol. (2006)

Mechanistic similarities between oxidation of hydroethidine by Fremy's salt and superoxide: stopped-flow optical and EPR studies

Authors:

Zielonka J, Zhao H, Xu Y, Kalyanaraman B.

Journal:

Free Radic Biol Med (2005): 853

Physical properties

Molecular weight	315.41
Solvent	DMSO

Spectral properties

Excitation (nm)	500
Emission (nm)	582

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200
CAS	104821-25-2

Instrument settings

Fluorescence microplate reader
Excitation	520 nm
Emission	600 nm
Cutoff	550 nm
Recommended plate	Solid black

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Page updated on October 8, 2025