NADPH Oxidases: Methods and Protocols

by Knaus, Ulla G.

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Title Statements

Full title :NADPH Oxidases : Methods and Protocols.

Summary

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

Edition Statements

Publication Date:2019.

Publication Statements

Publisher(s):Springer
Place of Publication:New York, NY
Language:eng
Publication Date:2019.

Notes

3.2.2 Expression and Purification of His-p67phox

The Physical Object

Description:1 online resource (708 pages)

Edition Identifiers

ISBN:9781493994243, 1493994247
DOI:10.1007/978-1-4939-9424-3.
oclc:on1122916040
LC Classification:QH301-705
Dewey Decimal:572.6

Intro; Preface; Contents; Contributors; Part I: The NADPH Oxidase Family: Overviews; Chapter 1: Intersecting Stories of the Phagocyte NADPH Oxidase and Chronic Granulomatous Disease; 1 Introduction; 2 The ``Respiratory Burst;́́ 3 Essential and Recurrent Role of Clinicians; 4 Cytochrome b558; 5 Cytoplasmic Components of the Phagocyte Oxidase; 6 Regulation of the Phagocyte NADPH Oxidase; 7 Summary; References; Chapter 2: Mammalian NADPH Oxidases; 1 Introduction; 2 The NOX Family Members; 2.1 NOX2; 2.2 NOX1; 2.3 NOX3; 2.4 NOX4; 2.5 NOX5; 2.6 DUOX1 and DUOX2; 3 Conclusion; References

Part II: Biochemistry and Model SystemsChapter 3: Enhanced Immunoaffinity Purification of Human Neutrophil Flavocytochrome B for Structure Determination by Electron ... ; 1 Introduction; 2 Materials; 2.1 Preparation of the mAb CS9 Affinity Purification Matrix; 2.2 Immunoaffinity Purification of Cytb; 2.3 Concentration and Desalting of Cytb; 2.4 Reconstitution of Cytb in 99% Phosphatidylcholine Vesicles by Dialysis; 2.5 Negative-Stain Transmission Electron Microscopy (TEM) of Detergent-Solubilized Cytb; 2.6 Cryoelectron Microscopy (CryoEM); 3 Methods

3.1 Preparation of the CS9 mAb Affinity Matrix3.2 Immuno…