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Detecting Reactive Oxygen Species

Overview

Reactive oxygen species are chemically active, oxygen-derived molecules capable of oxidizing other molecules. Because of their reactive nature, there are many deleterious effects associated with unchecked ROS production, including structural damage to DNA and other biological molecules. However, ROS can also be mediators of physiological signaling. There is accumulating evidence that ROS play significant roles in everything from activation of transcription factors to the mediation of inflammatory toxicity that kills foreign pathogens and defend the body.

In this video we will delve into the associations between ROS, metabolism and disease. After establishing their significance, we will discuss the principles and a protocol of a commonly used methodology for measuring ROS levels in cells: the use of non-fluorescent probes that become fluorescent upon oxidation. Lastly, we will review some current applications of this technique in cell biology research.

Procedure

Reactive oxygen species produced in cells have been implicated in tissue homeostasis, cellular aging, and disease states like cancer. As their name implies, these molecules arise from oxygen, which naturally exists as a stable, dioxygen molecule since all its electrons are paired. The addition of one unpaired electron renders it unstable, and leads to formation of the superoxide anion—a form of reactive oxygen species or ROS. Other than the superoxide anion, there are several types of reactive species with unpaired e

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Tags
Reactive Oxygen SpeciesTissue HomeostasisCellular AgingDisease StatesCancerStable Dioxygen MoleculeUnpaired ElectronSuperoxide AnionROSCell MetabolismFluorescent Probe AssayGeneralized ProtocolMitochondriaElectron Leakage

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0:00

Overview

1:15

Cell Metabolism, Disease and ROS

3:43

Principles of ROS Measurement Using a Fluorescent Probe

4:52

Generalized Protocol for Measuring ROS

5:52

Applications

8:16

Summary

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