Mitochondrial Hydroxyl Radical Detection Assay Kit (ab219931)

Overview

  • Product name

    Mitochondrial Hydroxyl Radical Detection Assay Kit
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Species reactivity

    Reacts with: Other species, Mammals
  • 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.

  • 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.

  • Platform

    Microplate reader, Fluorescence microscope

Properties

  • 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

  • Hydroxyl radical production in HeLa cells. HeLa cells (10cells/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.

  • 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.

Protocols

References

ab219931 has not yet been referenced specifically in any publications.

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