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Dihydroethidium (Hydroethidine)
Citations & References (366)
Invitrogen™

Dihydroethidium (Hydroethidine)

Free radical sensor

Have Questions?
Catalog NumberQuantity
D231071 mL
Catalog number D23107
Price (MXN)
-
Quantity:
1 mL

The superoxide indicator dihydroethdium, also called hydroethidine, exhibits blue-fluorescence in the cytosol until oxidized, where it intercalates within the cell's DNA, staining its nucleus a bright fluorescent red.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
For Use With (Application)Superoxide Indicator
No. of Tests1 mL
Quantity1 mL
Detection MethodFluorescence
Emission Wavelength Range606 nm
Excitation/Emission518/606 nm
Excitation Wavelength Range518 nm
Product TypeROS Indicator
Unit SizeEach

Frequently asked questions (FAQs)

I want to assay cells for reactive oxygen species using carboxy-H2DCFDA, but I want to do so with a plate reader instead of microscope. Will it work?

It has been done. The problem is that plate readers are less sensitive than microscopes, with far less signal-to-background difference. It is worth trying, but first optimize concentrations and loading times with control cells, use a plate with little to no autofluorescence, and possibly optimize the gain setting in order to get the best signal possible. But don't expect the same sensitivity, even with optimization.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I have GFP-transfected cells and need to label for reactive oxygen species. Can I use H2DCFDA?

This is not recommended as the two dyes overlap in the emission wavelength. There are other ROS reagents available in different wavelengths, such as CellROX Deep Red, which emits in the far-red range (665 nm), or dihydroethidium, which is emits in the visible red range (620 nm).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Citations & References (366)

Citations & References
Abstract
Authors:
Journal:
PubMed ID:10891486
In vivo evidence for microvascular oxidative stress in spontaneously hypertensive rats. Hydroethidine microfluorography.
Authors:Suzuki H, Swei A, Zweifach BW, Schmid-Schönbein GW
Journal:Hypertension
PubMed ID:7737720
The factors that predispose to the accelerated organ injury that accompanies the hypertensive syndrome have remained speculative and without a firm experimental basis. Indirect evidence has suggested that a key feature may be related to an enhanced oxygen radical production. The purpose of this study was to refine and use ... More
Evaluation of a flow cytometric fluorescence quenching assay of phagocytosis of sensitized sheep erythrocytes by polymorphonuclear leukocytes.
Authors:Van Amersfoort ES, Van Strijp JA
Journal:Cytometry
PubMed ID:7875036
A number of reports have been published describing phagocytosis assays for flow cytometric analysis. In some of these, the fluorescence quenching technique has been used to discriminate between adherent and ingested particles. In this report, we have evaluated the efficacy of a quantitative fluorescence quenching technique with crystal violet and ... More
The human salivary peptide histatin 5 exerts its antifungal activity through the formation of reactive oxygen species.
Authors:Helmerhorst EJ, Troxler RF, Oppenheim FG
Journal:Proc Natl Acad Sci U S A
PubMed ID:11717389
Previous studies have shown that the human salivary antifungal peptide histatin 5 is taken up by Candida albicans cells and associates intracellularly with mitochondria. The purpose of the present study was to investigate the biological consequence of this specific subcellular targeting. Histatin 5 inhibited respiration of isolated C. albicans mitochondria ... More
Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide.
Authors:Zhao H, Kalivendi S, Zhang H, Joseph J, Nithipatikom K, Vásquez-Vivar J, Kalyanaraman B
Journal:Free Radic Biol Med
PubMed ID:12757846
Hydroethidine (HE) or dihydroethidium (DHE), a redox-sensitive probe, has been widely used to detect intracellular superoxide anion. It is a common assumption that the reaction between superoxide and HE results in the formation of a two-electron oxidized product, ethidium (E+), which binds to DNA and leads to the enhancement of ... More
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