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 EVOS® Cell Imaging Systems
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 Cell Imaging
  Ready Probes® Reagents
  Cell Health: Detection
  Oxidative Stress
  Cell Proliferation
  Autophagy
  Ion Indicators
Oxidative Stress


Generation of reactive oxygen species (ROS) is inevitable for aerobic organisms and, in healthy cells, occurs at a controlled rate. Under conditions of oxidative stress, ROS production is dramatically increased, resulting in subsequent alteration of membrane lipids, proteins, and nucleic acids. Oxidative damage of these biomolecules is associated with aging as well as a variety of pathological events, including atherosclerosis, carcinogenesis, ischemia reperfusion injury, and neurodegenerative disorders.

 

 

We provide a variety of Molecular Probes® fluorescent tools to track different parameters in oxidative stress:

These tools use platforms such as fluorescence microscopy, flow cytometry, or microplate analysis.

 

Generalized oxidative stress

 are fluorogenic probes for measuring generalized oxidative stress in cells using conventional fluorescence microscopy, high content screening, microplate fluorometry, or flow cytometry. The dyes are nonfluorescent in a reduced state and fluoresce bright green, orange, or deep red upon oxidation. Some are formalin fixable for easy multiplexing with other fluorescent reagents including antibodies. Assays are simple and reliable with a sensitive and robust readout, and the reagent can be applied to cells in complete growth media. We provide CellROX® dyes and conventional probes for oxidative stress measurement.

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  Oxidative stress detection with CellROX® Green Reagent
General oxidative stress detection in U2OS cells with .

Lipid peroxidation

Lipid peroxidation is the oxidative degradation of lipids. ROS are the major initiators of lipid peroxidation, and membrane-bound polyunsaturated fatty acids like arachidonic acid and linoleic acid are their major targets. The byproducts of lipid peroxidation cause direct damage to cell membranes. They also form protein adducts, resulting in cell and tissue damage. Lipid peroxidation is implicated in many human diseases including diabetes and cardiovascular disease. We provide Molecular Probes® tools for live and fixed cells, with a choice of ratiometric or single-wavelength readout.

  Detection of lipid peroxidation with the Image-iT® Lipid Peroxidation Kit
Detection of lipid peroxidation with the.

Selective ROS detection

Superoxide, peroxyl radical, hydrogen peroxide, hydroxyl radical, and peroxynitrite are examples of ROS that react with nucleic acids, proteins, and lipids and result in cell and tissue damage. Certain ROS have been implicated in various human diseases including cancer, cardiovascular disease, neurodegenerative disease, and aging. We provide selective Molecular Probes® reagents for a variety of reactive species targeted to the cytosol or mitochondria and detected using imaging, flow cytometry, or microplate analysis.

  Detection of ROS with MitoSOX™ Red Superoxide Indicator
Detection of superoxide in live cells using .

Fluorescent protein–based redox sensors

Genetically encoded sensors provide an alternative to organic dyes for measuring redox potential or oxidative stress in live cells. roGFP is a fluorescent protein chimera that changes its excitation maximum from 488 nm to 400 nm based on a highly efficient redox relay. Using ratiometric detection of the emission at 515 nm, this shift can be used to measure H2O2 (Premo™ Cellular Hydrogen Peroxide H2O2Sensor) or redox potential (Premo™ Cellular Redox Sensor, Grx-1-roGFP)  in live cells in a reversible and reproducible assay. The fluorescent proteins can be multiplexed with organic dyes or other expressed proteins to provide additional biological context for the assay.

  Premo™ Cellular Redox Sensor
Detecting changes in cellular redox potential in U2OS cells using . 

Glutathione

Reduced glutathione, also known as GSH, is a major thiol bound to proteins. Protein thiols including GSH play an important role in determining the redox status of cells. Therefore, detection of GSH levels is a useful indication of redox potential and a cell’s ability to prevent oxidative stress. We provide Molecular Probes® intracellular probes for the sensitive detection and localization of thiols.

  Detection of glutathione with ThiolTracker™ Violet dye
Detection of GSH in U2OS cells with dye.


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