DHE (Dihydroethidium) Assay Kit - Reactive Oxygen Species (ab236206)
Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader, Flow cytometer
- Sample type: Adherent cells, Suspension cells
Overview
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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.
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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.
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Platform
Microplate reader, Flow cytometer
Properties
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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
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Example Data.
THP-1 cells (human monocytic leukemia cell line) treated with controls and stained using the protocol for suspension cells.
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Example Data.THP-1 cells (human monocytic leukemia cell line) treated with controls and stained using the protocol for suspension cells.
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Example Data.THP-1 cells (human monocytic leukemia cell line) 70k cells/well, treated with controls and stained using the protocol for suspension cells.
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Example Data.HepG2 cells (human liver hepatocellular carcinoma cell line) 60k cells/well, treated with controls and stained using the protocol for adherent cells.