Mitochondrial Superoxide Assay Kit (Fluorometric) (ab219943)

Overview

  • Product name

    Mitochondrial Superoxide Assay Kit (Fluorometric)
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Assay type

    Cell-based
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    Mitochondrial Superoxide Detection Kit (Fluorometric) (ab219943) is a sensitive fluorometric one-step assay to detect intracellular superoxide radical in live cells. The assay uses our MitoROS 580 dye: the dye is cell-permeable and selectively reacts with mitochondrial superoxide 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

    Mitochondria are major producers of cellular superoxide. The production of low to moderate levels of superoxide is critical for the proper regulation of many essential cellular processes including gene expression, signal transduction, and muscle adaptation to endurance exercise training. Uncontrolled mitochondrial superoxide production can trigger cellular oxidative damage that contributes to the pathogenesis of a wide variety of disorders including cancer, cardiovascular diseases, neurodegenerative diseases and aging. The detection of intracellular mitochondrial superoxide is of central importance to understanding proper cellular redox regulation and the impact of its dysregulation on various pathologies.

    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
    MitoROS™ 580 1 vial

Images

  • Superoxide production in HeLa cells. HeLa cells were seeded overnight (105 cells/well/100 µL) in a 96 well black wall/clear bottom plate. Left: cells were treated with 50 µM Antimycin A (AMA) at 37ºC for 30 minutes, then incubated with MitoROS 580 for 1 hour. Right: control HeLa cells were incubated with MitoROS580 at 37 ºC for 1 hour without treatment. The fluorescence signal was measured using fluorescence microscope with a TRITC filter.

  • Quantification of superoxide production in HeLa cells. HeLa cells were seeded overnight (105 cells/well/100 µL) in a 96 well black wall/clear bottom plate. Cells were left untreated (control) or treated with either pyocyanin (Pyo, 50 µM Pyocyanin) or antimycin A (AMA, 50 µM Antimycin A) at 37 ºC for 30 minutes. Cells were then incubated with MitoROS 580 at 37 ºC for 1 hour. The fluorescence signal was monitored at Ex/Em = 540/590 nm (cut off = 570 nm) with bottom read mode using a microplate reader.

Protocols

References

This product has been referenced in:

  • Nguyen TT  et al. Oxidative stress induced by Porphyromonas gingivalis lysate and nicotine in human periodontal ligament fibroblasts. Odontology N/A:N/A (2018). Read more (PubMed: 29959559) »
See 1 Publication for this product

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