Unit

UNIT 9.35 Detection of Intracellular Glutathione Using ThiolTracker Violet Stain and Fluorescence Microscopy

  1. Bhaskar S. Mandavilli,
  2. Michael S. Janes

Published Online: 1 JUL 2010

DOI: 10.1002/0471142956.cy0935s53

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Mandavilli, B. S. and Janes, M. S. 2010. Detection of Intracellular Glutathione Using ThiolTracker Violet Stain and Fluorescence Microscopy. Current Protocols in Cytometry. 53:9.35:9.35.1–9.35.8.

Author Information

  1. Invitrogen Corporation, Molecular Probes Labeling and Detection Technologies, Eugene, Oregon

Publication History

  1. Published Online: 1 JUL 2010
  2. Published Print: JUL 2010

Abstract

Glutathione plays an important role in protecting mammalian cells from oxidative stress and cell death. Because reduced glutathione (GSH) represents the large majority of intracellular free thiols, cell-permeant, thiol-reactive fluorescent probes represent potentially useful indicators of intracellular GSH. The ThiolTracker Violet stain (a registered trademark of Invitrogen) is a bright fluorescent probe that is highly reactive to thiols and can be used as a convenient and effective indicator of intracellular GSH and general redox status by a variety of detection modalities. While this probe has been validated in flow cytometry and microplate fluorimetry assays, the following method will describe details on the use of the ThiolTracker Violet dye in traditional fluorescence microscopy, as well as high-content imaging and analysis. Curr. Protoc. Cytom. 53:9.35.1-9.35.8. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • glutathione;
  • ThiolTracker Violet;
  • oxidative stress;
  • cell based assays;
  • high content imaging;
  • fluorescence microscopy