Search

  • Order Status
  • Quick Order
  • Promos
Carboxy-H2DCFDA (general oxidative stress indicator)
Citations & References (95)
Invitrogen™

Carboxy-H2DCFDA (general oxidative stress indicator)

General oxidative stress indicator

Have Questions?
Catalog NumberQuantity
C40025 mg
Catalog number C400
Price (EUR)
435,65
Online exclusive
474,00
Save 38,35 (8%)
Each
Quantity:
25 mg
Price (EUR)
435,65
Online exclusive
474,00
Save 38,35 (8%)
Each

6-carboxy-2',7'-dichlorodihydrofluorescein diacetate is a chemically reduced, acetylated form of fluorescein used as an indicator for reactive oxygen species (ROS) in cells. This nonfluorescent molecule is readily converted to a green-fluorescent form when the acetate groups are removed by intracellular esterases and oxidation (by the activity of ROS) occurs within the cell. We offer the carboxylated H2DCFDA analog (carboxy-H2DCFDA, Cat. No. C400), which has two negative charges at physiological pH, and its di(acetoxymethyl ester) (Cat. No. C2938). Upon cleavage of the acetate and ester groups by intracellular esterases and oxidation, both analogs form carboxydichlorofluorescein (Cat. No. C368), with additional negative charges that impede its leakage out of the cell.

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. Not for human or animal therapeutic or diagnostic use.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
For Use With (Application)ROS
Quantity25 mg
Detection MethodFluorescence
Emission Wavelength Range517 to 527 nm
Excitation Wavelength Range492 to 495 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.

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.

Citations & References (95)

Citations & References
Abstract
N-acetylcysteine augments the cellular redox changes and cytotoxic activity of internalized mycobacterium bovis in human bladder cancer cells.
Authors:Pook SH, Esuvaranathan K, Mahendran R
Journal:J Urol
PubMed ID:12131368
PURPOSE: We determined whether changes in cellular reactive oxygen species correlated with mycobacteria internalization and bladder cancer cell death. MATERIALS AND METHODS: Reactive oxygen species and thiols in RT112 and MGH bladder cancer cells were determined using the fluorescence probes 5-(and 6)-carboxy-2', 7' dichlorodihydrofluorescein diacetate and monobromobimane. Superoxide and nitrite ... More
The sulphonylurea glibenclamide inhibits multidrug resistance protein (MRP1) activity in human lung cancer cells.
Authors:Payen L, Delugin L, Courtois A, Trinquart Y, Guillouzo A, Fardel O
Journal:Br J Pharmacol
PubMed ID:11159731
'1. Glibenclamide, a sulphonylurea widely used for the treatment of non-insulin-dependent diabetes mellitus, has been shown to inhibit the activities of various ATP-binding cassette (ABC) transporters. In the present study, its effects towards multidrug resistance protein 1 (MRP1), an ABC efflux pump conferring multidrug resistance and handling organic anions, were ... More
Characterization of Vta1p, a class E Vps protein in Saccharomyces cerevisiae.
Authors:Shiflett SL, Ward DM, Huynh D, Vaughn MB, Simmons JC, Kaplan J
Journal:J Biol Chem
PubMed ID:14701806
'We identified VTA1 in a screen for mutations that result in altered vacuole morphology. Deletion of VTA1 resulted in delayed trafficking of the lipophilic dye FM4-64 to the vacuole and altered vacuolar morphology when cells were exposed to the dye 5-(and 6)-carboxy-2'',7''-dichlorofluorescein diacetate (CDCFDA). Deletion of class E vacuolar protein ... More
Mitochondrial localization of reactive oxygen species by dihydrofluorescein probes.
Authors:Diaz G, Liu S, Isola R, Diana A, Falchi AM
Journal:Histochem Cell Biol
PubMed ID:14574587
'Mitochondria are the main source of reactive oxygen species (ROS). The aim of this work was to verify the ROS generation in situ in HeLa cells exposed to prooxidants and antioxidants (menadione, tert-butyl hydroperoxide, antimycin A, vitamin E, N-acetyl-L-cysteine, and butylated hydroxytoluene) using the ROS-sensitive probes 6-carboxy-2'',7''-dichlorodihydrofluorescein diacetate di-acetomethyl ester ... More
The use of fluorescent probes to assess oxidative processes in isolated-perfused rat heart tissue.
Authors:Kehrer JP, Paraidathathu T
Journal:Free Radic Res Commun
PubMed ID:1505782
'The formation of reactive oxygen species (ROS) in intact heart tissue has been assessed by direct ESR measurements, and indirectly by the formation of characteristic tissue products and the protective effects of various antioxidants. The development of lipid soluble esters of compounds which can be trapped intracellularly after hydrolysis, and ... More
View all Carboxy-H2DCFDA (general oxidative stress indicator) citations