Mitochondrial Hydroxyl Radical Detection Assay Kit (ab219931) is a sensitive fluorometric one-step assay to detect intracellular hydroxyl radical (OH·) in live cells.
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- Antioxidants (Basel, Switzerland) 12:2023Curcumin and N-Acetylcysteine Nanocarriers Alone or Combined with Deferoxamine Target the Mitochondria and Protect against Neurotoxicity and Oxidative Stress in a Co-Culture Model of Parkinson's Disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLeah Mursaleen,Stefanie Ho Yi Chan,Brendon Noble,Satyanarayana Somavarapu,Mohammed Gulrez ZariwalaPubMed 36670992
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- Redox biology 37:1016982020Non-thermal dielectric-barrier discharge plasma induces reactive oxygen species by epigenetically modifying the expression of NADPH oxidase family genes in keratinocytes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKyoung Ah Kang et. al.PubMed 32863235
- Antioxidants (Basel, Switzerland) 9:2020N-Acetylcysteine Nanocarriers Protect against Oxidative Stress in a Cellular Model of Parkinson's Disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLeah Mursaleen et. al.PubMed 32660079
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Reactive species
- Mammals
Target data
What's included?
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Mitochondrial Hydroxyl Radical Detection Assay Kit (ab219931) is a sensitive fluorometric one-step assay to detect intracellular hydroxyl radical (OH·) in live cells.
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Reactive species
- Mammals
- Assay Platform
- Microplate reader, Fluorescence microscope
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
Mitochondrial Hydroxyl Radical Detection Assay Kit (ab219931) is a sensitive fluorometric one-step assay to detect intracellular hydroxyl radical (OH·) in live cells. The assay uses our OH580 probe: the probe is cell-permeable and selectively reacts with hydroxyl radical present in live cells to generate a red fluorescence signal that can be easily read at Ex/Em= 540/590 nm.
The assay can be performed within one hour and can be detected by fluorescence microscopy, microplate reader or high-content imaging. It can be easily adapted to use in 384-well microplate format.
The detection of intracellular hydroxyl radical is of central importance to understanding proper cellular redox regulation and the impact of its dysregulation on various pathologies. The hydroxyl radical (HO·) is one of the reactive oxygen species (ROS) highly reactive with other molecules to achieve stability. In general, hydroxyl radical is considered to be a harmful by-product of oxidative metabolism, which can cause molecular damage in living system. It shows an average lifetime of 10-9 s and can react with nearly every biomolecule such as nuclear DNA, mitochondrial DNA, proteins and membrane lipids.**Related products**Review the , or the full to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Mitochondrial hydroxyl radicals often abbreviated as 'OH radicals' are highly reactive species that derive from the reduction of oxygen. These radicals are produced in the mitochondria the energy powerhouses of the cell and are part of reactive oxygen species (ROS). The precise mass of individual hydroxyl radicals is not typically measured but these small molecules play a critical role in cellular functions. Mitochondrial ROS detection is an area of intense study due to its implications in understanding cellular oxidative stress.
- Biological function summary
The presence of hydroxyl radicals can lead to oxidative damage to vital macromolecules including DNA proteins and lipids. The mitochondria themselves are frequent sites of this damage. Hydroxyl radicals are not part of a specific molecular complex but interact with various other molecules within the cell initiating chain reactions that propagate oxidative damage. This kind of impairment is involved in the regulation of apoptosis and other cellular activities.
- Pathways
These radicals are central to the oxidative stress pathway. The antioxidant system often counters their effects by reducing the potential damage to cellular structures. Proteins such as superoxide dismutase (SOD) and catalase are related to this pathway and play an important role in mitigating the detrimental effects of oxidative stress. These pathways ensure a delicate balance between ROS production and elimination important for maintaining cellular homeostasis.
- Associated diseases and disorders
Mitochondrial hydroxyl radicals have been linked to neurodegenerative diseases such as Alzheimer's and Parkinson's disease. The unregulated production of these radicals can lead to significant cellular damage contributing to the progression of these conditions. Proteins like amyloid-beta in Alzheimer's disease and alpha-synuclein in Parkinson's disease connect to hydroxyl radicals through oxidative stress mechanisms where excessive ROS contributes to protein aggregation and neuronal damage. Understanding the interaction of hydroxyl radicals with these proteins offers insights into potential therapeutic interventions.
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2 product images
Fluorescence Microscopy - Mitochondrial Hydroxyl Radical Detection Assay Kit (ab219931)
Hydroxyl radical production in HeLa cells. HeLa cells (105 cells/well/100 μL) were incubated with OH580 working solution at 37°C for 1 hour, then washed once with HHBS. Left: cells were treated with Fenton Reaction (10 μM CuCl2 and 100 μM H2O2) in 1X HBSS buffer at 37°C for 1 hour. Right: control HeLa cells in 1X HBSS buffer without treatment. After 3 washes with HHBS, HeLa cells were measured using a fluorescence microscope with a TRITC filter set (Red). Cell nuclei were stained with Hoechst 33342.
Fluorescence Microscopy - Mitochondrial Hydroxyl Radical Detection Assay Kit (ab219931)
Intracellular hydroxyl radical in RAW 264.7 cells. Cells were incubated with OH580 working solution at 37°C for 1 hour, then washed once with HHBS. Cells were then incubated without or with PMA (phorbol 12-myristate 13-acetate, 10-500 ng/mL) in growth medium at 37°C for 4 hours. After washing 3 times with HHBS, HeLa cells were measured using a fluorescence microscope with a TRITC filter set.
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