Shop All Reactive Oxygen & Nitrogen Species

RedoxSensor™ Red CC-1 - Special Packaging Invitrogen™

RedoxSensor Red CC-1 is a unique probe whose fluorescence localization appears to be based on a cell's cytosolic redox potential. Upon passively entering live cells, the stain is either oxidized in the cytosol to a red-fluorescent product (excitation/emission maxima ~540/600 nm), which then accumulates in mitochondria, or the probe is transported to lysosomes, where it is subsequently oxidized. The differential distribution of the oxidized product appears to depend on the redox potential of the cytosol.

Di(Acetoxymethyl Ester) (6-Carboxy-2',7'-Dichlorodihydrofluorescein Diacetate) Invitrogen™

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, C400), which has two negative charges at physiological pH, and its di(acetoxymethyl ester) (C2938). Upon cleavage of the acetate and ester groups by intracellular esterases and oxidation, both analogs form carboxydichlorofluorescein (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.

Image-IT™ LIVE Green Reactive Oxygen Species Detection Kit, for microscopy Invitrogen™

The Image-iT™ LIVE Green Reactive Oxygen Species (ROS) Detection Kit provides the key reagents necessary for the detection of ROS in live cells. The assay is based on 5-(and-6)-carboxy-2',7'-dichlorodihydrofluorescein diacetate (carboxy-H2DCFDA), a reliable fluorogenic marker for ROS in live cells. In addition to carboxy-H2DCFDA, the kit provides the common inducer of ROS production tert-butyl hydroperoxide (TBHP), as a positive control and the blue-fluorescent, cell-permeant nucleic acid stain Hoechst 33342.

Singlet Oxygen Sensor Green - Special Packaging Invitrogen™

Singlet Oxygen Sensor Green is a detection reagent that is highly selective for singlet oxygen. Unlike other available fluorescent and chemiluminescent singlet oxygen detection reagents, Singlet Oxygen Sensor Green does not show any appreciable response to hydroxyl radical or superoxide. This indicator initially exhibits weak blue fluorescence, but in the presence of singlet oxygen, it emits a green fluorescence (excitation/emission maxima ~504/525 nm) similar to that of fluorescein. Singlet Oxygen Sensor Green is cell-impermeant.

Dihydrorhodamine 123 Invitrogen™

Dihydrorhodamine 123 is an uncharged and nonfluorescent reactive oxygen species (ROS) indicator that can passively diffuse across membranes where it is oxidized to cationic rhodamine 123 which localizes in the mitochondria and exhibits green fluorescence.

OxyBURST™ Green H2DCFDA, SE Invitrogen™

OxyBURST® Green H2DCFDA succinimidyl ester (2',7'-dichlorodihydrofluorescein diacetate, SE) is an amine-reactive assay reagent that can be used to prepare oxidation-sensitive conjugates of a wide variety of biomolecules and particles, including antibodies, antigens, peptides, proteins, dextrans, bacteria, yeast and polystyrene microspheres. Following conjugation to amines, the two acetates of OxyBURST® Green H2DCFDA can be removed by treatment with hydroxylamine at neutral pH to yield the dihydrofluorescein conjugate. OxyBURST® Green H2DCFDA conjugates are nonfluorescent until they are oxidized to the corresponding fluorescein derivatives.

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.

DAF-FM Diacetate (4-Amino-5-Methylamino-2',7'-Difluorofluorescein Diacetate) Invitrogen™

DAF-FM is a reagent that is used to detect and quantify low concentrations of nitric oxide (NO). It is essentially nonfluorescent until it reacts with NO to form a fluorescent benzotriazole. DAF-FM fluorescence can be detected by any instrument that can detect fluorescein, including flow cytometers, microscopes, fluorescent microplate readers and fluorometers.

DAF-FM Specifications:
• Ex/Em of DAF-FM: ~495/515 nm
• Lyophilized product should be dissolved using DMSO and then added to an aqueous buffer to create a working solution
• DAF-FM diacetate is cell permeant and passively diffuses across cellular membranes; once inside the cell, it is converted to a cell-impermeant form
• Buffers containing bovine serum albumin (BSA) and phenol red may affect the fluorescence and should be used with caution
• The fluorescence quantum yield of DAF-FM is ~0.005, but increases about 160-fold, to ~0.81, after reacting with NO


Applications of DAF-FM:
• Assessment of NO production in transaldolase-deficient lymphoblasts by flow cytometry
• Detection of NO accumulation in embryonic cortical neurons following neurotrophin stimulation
in vivo imaging of NO in zebrafish
• Intravital microscopic detection of NO generation associated with angiogenesis in mice
• Quantitation of ATP-induced NO release in rabbit platelets

Important Advantages of DAF-FM Over DAF-2
The spectra of the NO adduct of DAF-FM are independent of pH above pH 5.5. Also, the NO adduct of DAF-FM is significantly more photostable than that of DAF-2, which means additional time for image capture. Finally, DAF-FM is a more sensitive reagent for NO than is DAF-2 (NO detection limit for DAF-FM ~3 nM versus ~5 nM for DAF-2).

Probes for Nitric Oxide Research
We offer several products for the detection of reactive oxygen species (ROS) and nitric oxide (NO). Review these products in Probes for Nitric Oxide Research—Section 18.3 in the Molecular Probes® Handbook.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.

Image-iT™ Lipid Peroxidation Kit, for live cell analysis Invitrogen™

The Image-iT® Lipid Peroxidation Kit enables the detection of lipid peroxidation in live cells through oxidation of BODIPY® 581/591 C11 reagent. This reagent localizes to membranes throughout live cells and upon oxidation by lipid hydroperoxides, displays a shift in peak fluorescence emission from ~590 nm to ~510 nm. Fluorescence from live cells shifts from red to green, providing a ratiometric indication of lipid peroxidation compatible with traditional and high content microscopy, as well as flow cytometry. This kit also includes cumene hydroperoxide as a positive control compound to induce lipid peroxidation in cells.

Additional Information:

• The reagent is provided as a stable, ready-to-use DMSO solution with a simple protocol compatible with standard workflows in fluorescence microscopy
• The reagent is provided as five single-use vials containing sufficient total reagent for five 96-well plates or 100 coverslips

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

Total Reactive Oxygen Species (ROS) Assay Kit 520 nm Invitrogen™

The Total Reactive Oxygen Species (ROS) Assay Kit 520 nm contains the necessary regent and buffer for identifying ROS in cells by flow cytometry in the FITC channel. Reactive oxygen species are chemically reactive oxygen-containing molecules that are generated as a natural byproduct of the oxygen metabolism. ROS are constantly generated and eliminated under normal physiologic conditions and have important functions in cell signaling, homeostasis, and clearance of microbial infections. During times of environmental stress such as exposure to UV or heat, or during infection, ROS levels can increase dramatically leading to oxidative stress. Oxidative stress can result in damage to cellular proteins, lipids, and DNA, and has been linked to cardiovascular disease, cancer, diabetes, inflammation, and aging. Examples of ROS include superoxide ions and peroxides.

Reported Application
Flow Cytometric Analysis

Fc OxyBURST™ Green Assay Reagent Invitrogen™

The Fc OxyBURST® Green assay reagent comprises bovine serum albumin (BSA) that has been covalently linked to dichlorodihydrofluorescein (H2DCF) and then complexed with a purified rabbit polyclonal anti-BSA antibody. When this immune complex binds to Fc receptors, the nonfluorescent H2DCF molecules are internalized within the phagovacuoloe and subsequently oxidized to green-fluorescent 2''7'-dichlorofluorescein.

CellROX™ Orange Reagent, for oxidative stress detection Invitrogen™

CellROX® Orange Reagent is a novel fluorogenic probe for measuring oxidative stress in live cells. This cell-permeant dye is non-fluorescent while in a reduced state and exhibits bright orange fluorescence upon oxidation by reactive oxygen species (ROS), with absorption/emission maxima of ~ 545/565 nm. While this reagent is not formaldehyde-fixable, it can be multiplexed with other live cell-compatible probes such as GFP, CellEvent® Caspase 3/7 Green reagent, Image-iT® DEAD Green, Green and Deep Red MitoTracker® dyes, and NucBlue™ reagent.

CellROX® Orange Reagent is compatible with various platforms, such as traditional fluorescence microscopy, high content screening (HCS), and flow cytometery. This reagent is also compatible with the Tali™ Image-based Cytometer.

In addition, CellROX® Orange Reagent is:

• Compatible with other live cell probes, including GFP, and may be added directly to serum-containing growth media
• Provided as a stable, ready-to-use DMSO solution with a simple protocol compatible with standard workflows in fluorescence microscopy
• Provided as five single-use vials containing enough total reagent for five 96-well plates or 100 coverslips

Oxidative stress results from an imbalance between the production of reactive oxygen species (ROS) and the ability of cells to scavenge them. ROS play an important role in the progression of several diseases, including inflammation, atherosclerosis, aging, and age-related degenerative disorders.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

Glutathione Ethyl Ester, Biotin Amide (BioGEE) (Glutathiolation Detection Reagent) - Special Packaging Invitrogen™

Biotinylated glutathione ethyl ester (BioGEE) is a cell-permeant, biotinylated glutathione analog for the detection of glutathiolation. Under conditions of oxidative stress, cells may transiently incorporate glutathione into proteins. Stressed cells incubated in BioGEE will also incorporate this biotinylated glutathione derivative into proteins, facilitating the identification of oxidation-sensitive proteins. Once these cells are fixed and permeabilized, you can detect glutathiolation levels with a fluorescent streptavidin conjugate using either flow cytometry or fluorescence microscopy. Proteins glutathiolated with BioGEE can also be extracted and analyzed by mass spectrometry or by Western blotting methods in conjunction with fluorophore- or enzyme-labeled streptavidin conjugates.

CM-H2DCFDA (General Oxidative Stress Indicator) Invitrogen™

CM-H2DCFDA is a chloromethyl derivative of H2DCFDA, useful as an indicator for reactive oxygen species (ROS) in cells. This indicator exhibits much better retention in live cells than H2DCFDA. CM-H2DCFDA passively diffuses into cells, where its acetate groups are cleaved by intracellular esterases and its thiol-reactive chloromethyl group reacts with intracellular glutathione and other thiols. Subsequent oxidation yields a fluorescent adduct that is trapped inside the cell, thus facilitating long-term studies.

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.

BODIPY™ 581/591 C11 (Lipid Peroxidation Sensor) Invitrogen™

BODIPY® 581/591 undecanoic acid can be used to detect reactive oxygen species (ROS) in cells and membranes. Oxidation of the polyunsaturated butadienyl portion of the dye results in a shift of the fluorescence emission peak from ~590 nm to ~510 nm.

HPF (Hydroxyl Radical and Peroxynitrite Sensor) Invitrogen™

Aminophenyl fluorescein (APF, A36003) and hydroxyphenyl fluorescein (HPF), two reactive oxygen species (ROS) indicators developed by T. Nagano, offer greater specificity and stability than dichlorodihydrofluorescein diacetate (H2DCFDA, D399). APF and HPF show limited non-selective reactivity and relatively high resistance to light-induced oxidation. Both of these new fluorescein derivatives are nonfluorescent until they react with the hydroxyl radical or peroxynitrite anion; APF will also react with the hypochlorite anion. Together, these two ROS indicators can selectively detect hypochlorite anion. Upon oxidation, both APF and HPF exhibit bright green fluorescence (excitation/ emission maxima ~490/515 nm), making them compatible with any instrument that can detect fluorescein.
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