Mitochondrial Hydroxyl Radical Detection Assay Kit (ab219931)
Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader, Fluorescence microscope
- Sample type: Adherent cells, Suspension cells
Overview
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Product name
Mitochondrial Hydroxyl Radical Detection Assay Kit -
Detection method
Fluorescent -
Sample type
Adherent cells, Suspension cells -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
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.
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Notes
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 oxidative stress marker and assay guide, or the full metabolism assay guide 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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Platform
Microplate reader, Fluorescence microscope
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 200 tests Assay Buffer 1 x 50ml DMSO 1 x 100µl OH580 1 vial
Images
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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 37ºC 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 37ºC 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.
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Mitochondrial Hydroxyl Radical Detection Assay Kit (ab219931)Intracellular hydroxyl radical in RAW 264.7 cells. Cells were incubated with OH580 working solution at 37ºC 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 37ºC for 4 hours. After washing 3 times with HHBS, HeLa cells were measured using a fluorescence microscope with a TRITC filter set.
