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NADPH Oxidases

Methods and Protocols

  • Book
  • © 2019

Overview

Editors:
  1. Ulla G. Knaus

    1. Conway Institute, School of Medicine, University College Dublin, Dublin, Ireland

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  2. Thomas L. Leto

    1. Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, USA

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    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 1982)

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Table of contents (39 protocols)

  1. Front Matter

    Pages i-xv
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  2. The NADPH Oxidase Family: Overviews

    1. Front Matter

      Pages 1-1
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    2. Intersecting Stories of the Phagocyte NADPH Oxidase and Chronic Granulomatous Disease

      • William M. Nauseef, Robert A. Clark
      Pages 3-16

    3. Mammalian NADPH Oxidases

      • Hélène Buvelot, Vincent Jaquet, Karl-Heinz Krause
      Pages 17-36

  3. Biochemistry and Model Systems

    1. Front Matter

      Pages 37-37
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    2. Enhanced Immunoaffinity Purification of Human Neutrophil Flavocytochrome B for Structure Determination by Electron Microscopy

      • Algirdas J. Jesaitis, Marcia Riesselman, Ross M. Taylor, Susan Brumfield
      Pages 39-59

    3. Purification and Characterization of DUOX Peroxidase Homology Domains (PHDs)

      • Jennifer L. Meitzler
      Pages 61-74

    4. A Close-Up View of the Impact of Arachidonic Acid on the Phagocyte NADPH Oxidase

      • Tania Bizouarn, Hager Souabni, Xavier Serfaty, Aicha Bouraoui, Rawand Masoud, Gilda Karimi et al.
      Pages 75-101

    5. NOX5 Cell-Free Assay for the High-Throughput Screening of Small Molecules

      • Fiona Augsburger, Delphine Rasti, Yves Cambet, Vincent Jaquet
      Pages 103-111

    6. Spectroscopy of NOX Protein Family Members

      • Yoko Nakano, William M. Nauseef
      Pages 113-120

    7. Soluble Regulatory Proteins for Activation of NOX Family NADPH Oxidases

      • Hideki Sumimoto, Reiko Minakami, Kei Miyano
      Pages 121-137

    8. Insights into the NOX NADPH Oxidases Using Heterologous Whole Cell Assays

      • Mary C. Dinauer
      Pages 139-151

    9. The X-CGD PLB-985 Cell Model for NOX2 Structure-Function Analysis

      • Sylvain Beaumel, Marie José Stasia
      Pages 153-171

    10. Functional Characterization of DUOX Enzymes in Reconstituted Cell Models

      • Agnieszka Korzeniowska, Ágnes P. Donkó, Stanislas Morand, Thomas L. Leto
      Pages 173-190

    11. Guidelines for the Detection of NADPH Oxidases by Immunoblot and RT-qPCR

      • Becky A. Diebold, S. Garrett Wilder, Xavier De Deken, Jennifer L. Meitzler, James H. Doroshow, James W. McCoy et al.
      Pages 191-229

  4. Detection of ROS

    1. Front Matter

      Pages 231-231
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    2. Methods for Detection of NOX-Derived Superoxide Radical Anion and Hydrogen Peroxide in Cells

      • Fiona Augsburger, Aleksandra Filippova, Vincent Jaquet
      Pages 233-241

    3. HPLC-Based Monitoring of Oxidation of Hydroethidine for the Detection of NADPH Oxidase-Derived Superoxide Radical Anion

      • Jacek Zielonka, Monika Zielonka, Balaraman Kalyanaraman
      Pages 243-258

    4. Visualization of Intracellular Hydrogen Peroxide with the Genetically Encoded Fluorescent Probe HyPer in NIH-3T3 Cells

      • Yulia G. Ermakova, Nataliya M. Mishina, Carsten Schultz, Vsevolod V. Belousov
      Pages 259-274

    5. Imaging Intracellular H2O2 with the Genetically Encoded PerFRET and OxyFRET Probes

      • Balázs Enyedi, Miklós Geiszt
      Pages 275-282
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Keywords

About this book

This detailed volume explores the NADPH oxidase family of enzymes in human physiology and genetic disease, in which early discoveries represent prime examples of the finest translational “from bed to bench and back” studies. Methods are included for testing assembly and function of multicomponent oxidase complexes and for analyzing reactive oxygen species (ROS) generation in different systems by various means, while addressing pitfalls of ROS probes currently being used, as well as protocols on NADPH oxidase regulation and their function in cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and cutting-edge, NADPH Oxidases: Methods and Protocols will aid researchers working with the NOX/DUOX family in continuing and expanding upon their vital research.

Editors and Affiliations

  • Conway Institute, School of Medicine, University College Dublin, Dublin, Ireland

    Ulla G. Knaus

  • Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, USA

    Thomas L. Leto

Bibliographic Information

  • Book Title: NADPH Oxidases

  • Book Subtitle: Methods and Protocols

  • Editors: Ulla G. Knaus, Thomas L. Leto

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-9424-3

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2019

  • Hardcover ISBN: 978-1-4939-9423-6Published: 07 June 2019

  • Softcover ISBN: 978-1-4939-9426-7Published: 14 August 2020

  • eBook ISBN: 978-1-4939-9424-3Published: 06 June 2019

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XV, 718

  • Number of Illustrations: 44 b/w illustrations, 111 illustrations in colour

  • Topics: Enzymology, Human Physiology, Human Genetics

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