Product ID: M1241

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This reduced form of ethidine shows a blue fluorescence (absorption/emission: 355/420nm) in cell cytoplasm until oxidization to form ethidium which becomes red fluorescent (absorption/emission: 518/605 nm) upon DNA intercalation.

Dihydroethidium (also called hydroethidium or hydroethidine) is the chemically reduced form of the commonly used DNA intercalating dye ethidium bromide (B-ring reduction). This reduced dye is therefore very useful for detection of oxidative activities in viable cells, including respiratory burst in phagocytes, superoxide generation in mitochondria or as a vital stain in flow cytometry for imaging and analysis of intact cells. It has also been shown to exhibit increased fluorescence in various models of apoptosis. Only once it is internalized and dehydrogenated (oxidized) to ethidium, can it intercalate into DNA. Because of their compromised membranes, only dead cells are typically labeled by ethidium bromide when it selectively binds to DNA. But dihydroethidium is a neutral probe and is able to penetrate the cell membrane of live cells, staining their cytoplasm blue as well as the chromatin/nucleus of living cells red. Dihydroethidium typically exhibits a uniform labeling of cells within 30-40 minutes. Cell lysis of dihydroethidium-labeled cells can best be accomplished using Triton X-100 containing buffers. In addition, dihydroethidium has been incorporated into high-throughput assays to measure the effect of secondary agents or drugs on ROS activity in a live cell format.

Technical Data
SKU M1241
CAS Number 104821-25-2
Unit Size 10mg
Alternative Names hydroethidium, hydroethidine, 3,8-Diamino-5,6-dihydro-5-ethyl-6-phenylphenanthridine, 2,7-Diamino-10-ethyl-9-phenyl-9,10-dihydrophenanthridine
Absorption 392nm
Emission Wavelength 410
Detection Method Fluorescence
Molecular Formula C₂₁H₂₁N₃
Molecular Weight 315.42
Soluble In DMF, DMSO, CH3CN, MeOH
Storage Conditions -20C, Desiccated, Protect From Light

References and Citations


    Breton CS, Aubry D, Ginet V, Puyal J, Heulot M, Widmann C, Duchosal MA, Nahimana A (2015) "Combinative effects of β-Lapachone and APO866 on pancreatic cancer cell death through reactive oxygen species production and PARP-1 activation" Biochimie, 116: 141-153
  • Ginet V, Puyal J, Rummel C, Aubry D, Breton C, Cloux AJ, Majjigapu SR, Sordat B, Vogel P, Bruzzone S, Nencioni A, Duchosal MA, Nahimana A. (2014) "A critical role of autophagy in antileukemia/lymphoma effects of APO866, an inhibitor of NAD biosynthesis." Autophagy. 10(4):603-17.
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  • Saiki I, Bucana CD, Tsao JY, Fidler IJ. (1986) "Quantitative fluorescent microassay for identification of antiproliferative compounds." J Natl Cancer Inst 77(6): 1235-1240.
  • Frey T. (1997) "Correlated flow cytometric analysis of terminal events in apoptosis reveals the absence of some changes in some model systems." Cytometry 28(3): 253-263.
  • Budd SL. (1997) "Mitochondrial membrane potential and hydroethidine-monitored superoxide generation in cultured cerebellar granule cells." FEBS Lett 415(1): 21-24.
  • Zuo L, Christofi FL, Wright VP, Bao S, Clanton TL. (2004) "Lipoxygenase-dependent superoxide release in skeletal muscle." J Appl Physiol 97: 661-8.
  • King A, Gottlieb E, Brooks DG, Murphy MP, Dunaief JL. (2004) "Mitochondria-derived reactive oxygen species mediate blue light-induced death of retinal pigment epithelial cells." Photochem Photobiol 79: 470-5.
  • Nielsen HG, Hagberg IA, Lyberg T. (2004) "Marathon running leads to partial exhaustion of ROS-generating capacity in leukocytes." Med Sci Sports Exerc 36: 68-73.
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