DHE (Dihydroethidium) Assay Kit - Reactive Oxygen Species (ab236206)

Overview

  • Product name

    DHE (Dihydroethidium) Assay Kit - Reactive Oxygen Species
  • Detection method

    Fluorescent
  • Sample type

    Adherent cells, Suspension cells
  • Species reactivity

    Reacts with: Human
  • Product overview

    DHE (Dihydroethidium) Assay Kit - Reactive Oxygen Species (ab236206) measures ROS directly in live cells. This kit uses DHE as a fluorescent probe for the detection of ROS generation and is specific for superoxide and hydrogen peroxide. Antimycin A, an inhibitor of complex III of the mitochondrial electron transport chain, is included as a positive control for ROS generation. N-acetyl Cysteine is included as an anti-oxidant control.


    ROS generation is represented as total DHE fluorescence. It is important that positive and negative controls be included in every experiment for all cell types tested. Data can be expressed as total fluorescence, geometric mean fluorescence (flow cytometry) or as % or controls.

  • Notes

    DHE has been shown to be oxidized by superoxide to form 2-hydroxyethidium (2-OH-E+) (ex 500-530 nm/em 590-620 nm) or by non-specific oxidation by other sources of reactive oxygen species (ROS) to form ethidium (E+) (ex 480 nm/em 576 nm). Given their narrow spectral range, distinguishing between the two species using filter-based optical systems is often difficult and can lead to misreporting of the species of ROS being generated.

  • Platform

    Microplate reader, Flow cytometer

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 96 tests
    Antimycin A Assay Reagent 1 vial
    Cell-Based Assay Buffer Tablet 1 tablet
    Dihydroethidium Assay Reagent 2 x 20µl
    N-acetyl Cysteine Assay Reagent 2 x 10mg

Images

  • THP-1 cells (human monocytic leukemia cell line) treated with controls and stained using the protocol for suspension cells.

  • THP-1 cells (human monocytic leukemia cell line) treated with controls and stained using the protocol for suspension cells.

  • THP-1 cells (human monocytic leukemia cell line) 70k cells/well, treated with controls and stained using the protocol for suspension cells.

  • HepG2 cells (human liver hepatocellular carcinoma cell line) 60k cells/well, treated with controls and stained using the protocol for adherent cells.

Protocols

References

ab236206 has not yet been referenced specifically in any publications.

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