H2DCFDA (H2-DCF, DCF)
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H2DCFDA (H2-DCF, DCF)
Citations & References (1016)
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H2DCFDA (H2-DCF, DCF)

The cell-permeant 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) (also known as dichlorofluores cin diacetate) is a chemically reduced form of fluorescein used asRead more
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Catalog NumberQuantity
D399100 mg
Catalog number D399
Price (CNY)
2,093.00
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Ends: 31-Dec-2025
2,818.00
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Quantity:
100 mg
Price (CNY)
2,093.00
飞享价
Ends: 31-Dec-2025
2,818.00
Save 725.00 (26%)
Each
Add to cart
The cell-permeant 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) (also known as dichlorofluores cin diacetate) is a chemically reduced form of fluorescein used as an indicator for reactive oxygen species (ROS) in cells, for example to detect the generation of reactive oxygen intermediates in neutrophils and macrophages. Upon cleavage of the acetate groups by intracellular esterases and oxidation, the nonfluorescent H2DCFDA is converted to the highly fluorescent 2',7'-dichlorofluorescein (DCF). Versions of this indicator that have better cellular retention are available (C400, C2938).

ROS Indicator Specifications:

• Ex/Em: ∼492–495/517–527 nm
• Product is air sensitive and should be stored under dry argon or nitrogen
• Product may be dissolved in DMSO, DMF, or ethanol for use
• Indicator is cell permeant (cell loading protocols are available in the literature)
• Fluorescence can be monitored using a flow cytometer, fluorometer, microplate reader, or fluorescence microscope, using excitation sources and filters appropriate for fluorescein

Find More ROS Indicators
We offer an assortment of Molecular Probes™ products for the generation of reactive oxygen species (ROS), including singlet oxygen, superoxide, hydroxyl radical and various peroxide and hydroperoxides, as well as for their fluorometric detection in solution. Review Generating and Detecting Reactive Oxygen Species—Section 18.2 in the Molecular Probes™ Handbook for more information on these products.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Quantity100 mg
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.

I labeled my cell with CM-H2DCFDA for reactive oxygen detection, but upon illuminating the cell there is a significant increase in fluorescence in the control cells. Why?

If the cell is overloaded with dye, the high intracellular concentration of the dye may lead to dye-dye quenching. Upon illumination, photobleaching will occur, which will reduce the dye-dye quenching and actually increase the fluorescence (for a while, but then it will start decreasing). To solve the problem, reduce the concentration and incubation time, and try a range of incubation times and concentrations.

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

I need a formaldehyde-fixable reactive oxygen species detection assay. Is H2 DCFDA fixable?

H2DCFDA and similar derivatives are not fixable. The same goes for dihydroethidium and dihydrorhodamine. However, CellROX Deep Red and CellROX Green are retained for a limited time upon fixation with formaldehyde. CellROX Green may be retained upon subsequent Triton X-100 permeabilization. Avoid the use of any acetone or alcohol-based fixatives or fixatives that include alcohol, such as formalin.

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

What is the recommended storage condition/shelf-life for H2DCFDA (H2-DCF, DCF) (Cat. No. D399)?

We guarantee stability of the product for a period of 3 months when stored as recommended in the freezer (-5 to -30 degrees C).

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

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Citations & References (1016)

Citations & References
Abstract
Nitric oxide protects cultured rat hepatocytes from tumor necrosis factor-alpha-induced apoptosis by inducing heat shock protein 70 expression.
Authors:Kim YM,de Vera ME,Watkins SC,Billiar TR
Journal:The Journal of biological chemistry
PubMed ID:8995451
Differing effects of copper,zinc superoxide dismutase overexpression on neurotoxicity elicited by nitric oxide, reactive oxygen species, and excitotoxins.
Authors:Ying W,Anderson CM,Chen Y,Stein BA,Fahlman CS,Copin JC,Chan PH,Swanson RA
Journal:Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism
PubMed ID:10698074
Authors:
Journal:
PubMed ID:11282963
Imaging of hydroperoxides in a rat glomerulus stimulated by puromycin aminonucleoside.
Authors:Aoyagi K,Akiyama K,Tomida C,Gotoh M,Hirayama A,Takemura K,Ueda A,Nagase S,Koyama A,Narita M
Journal:Kidney international. Supplement
PubMed ID:10412763
Cellular alterations produced by the experimental increase in intracellular calcium and the nature of protective effects from pretreatment with nimodipine.
Authors:Danks AM,Hammond DN,Wainer BH,Van Buskirk RG,Isaacson RL
Journal:Brain research. Molecular brain research
PubMed ID:1334195
The immortalized septal cell line, SN56 B5 G4, generated by the fusion of mouse septal area cells and neuroblastoma cells, was used to determine if nimodipine, an antagonist of voltage sensitive calcium 'L' channels, might act in a neuroprotective fashion when intracellular calcium levels were raised by incubation in ouabain ... More
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