Mitochondrial Hydroxyl Radical Detection Assay Kit
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(7 Publications)
- Fluorescence Microscopy
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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
Supplier Data
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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Product details
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.
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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.
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Publications (7)
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Cell death & disease 16:467 PubMed40592821
2025
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Frontiers in molecular biosciences 11:1460987 PubMed39297074
2024
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Antioxidants (Basel, Switzerland) 12: PubMed36670992
2023
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Small (Weinheim an der Bergstrasse, Germany) 18:e2106570 PubMed35263020
2022
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Antioxidants (Basel, Switzerland) 10: PubMed34073115
2021
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Redox biology 37:101698 PubMed32863235
2020
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Antioxidants (Basel, Switzerland) 9: PubMed32660079
2020
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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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