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Detection of Total Reactive Oxygen Species in Adherent Cells by 2′,7′-Dichlorodihydrofluorescein Diacetate Staining
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Biochemie
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JoVE Journal Biochemie
Detection of Total Reactive Oxygen Species in Adherent Cells by 2′,7′-Dichlorodihydrofluorescein Diacetate Staining

Detection of Total Reactive Oxygen Species in Adherent Cells by 2′,7′-Dichlorodihydrofluorescein Diacetate Staining

DOI:
10.3791/60682-v

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04:53 min

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June 23, 2020

•

,
1Department of Biochemistry and Molecular Biology,University of New Mexico

Kapitel

  • 00:04Introduction
  • 00:32Cell Seeding and DCFH-DA Solution Preparation
  • 01:32DCFH-DA Staining
  • 02:12Image Acquisition and Intensity Measurement
  • 03:29Results: Iron Treatment Increases ROS in Colorectal Cancer Cells
  • 04:07Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

Here, we present a protocol to detect total cellular reactive oxygen species (ROS) using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). This method can visualize cellular ROS localization in adherent cells with a fluorescence microscope and quantify ROS intensity with a fluorescence plate reader. This protocol is simple, efficient and cost-effective.

Tags

Reactive Oxygen SpeciesDCFH-DA StainingAdherent CellsFluorescence MicroscopyFluorescence Plate ReaderOxidative StressHCT116 Colorectal Cancer CellsFerrous SulfateDoxorubicinRIPA BufferBradford AssayFluorescence Intensity Quantification

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