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NADPH Oxidases pp 517-528 | Cite as

Neutrophil Extracellular Traps

  • Balázs RadaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1982)

Abstract

Neutrophil extracellular traps (NETs) are made of a network of extracellular strings of DNA that bind pathogenic microbes. Histones and several neutrophil granule proteins associated with the DNA framework damage entrapped microorganisms. Reactive oxygen species generated by the neutrophil NADPH oxidase have been shown to be essential to mediate NET release by several stimuli including numerous pathogenic bacteria. Although several methods have been used in the literature to detect NETs in vitro and in vivo, a consensus is urgently needed on the field to standardize the best NET-specific assays. In this chapter, two methods are described in details that can be used to detect NETs and to distinguish them from other mechanisms of neutrophil cell death. While NET-specific, these assays are also relatively simple and straightforward enabling their potential use by a wide audience.

Key words

NETs Neutrophil NADPH oxidase MPO-DNA complex NE-DNA complex Citrullinated histone Peptidylarginine deiminase 4 Nuclear decondensation 

Notes

Acknowledgments

This work was supported by the NIH grant 1 R01 HL136707-01A1 awarded to B. Rada. I thank previous graduate students Dae-goon Yoo, Madison Floyd, and Payel Sil, for taking the immunofluorescence images presented here and for optimizing and recording detailed protocols that helped writing the “Notes” section of this book chapter.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.College of Veterinary Medicine, Department of Infectious DiseasesUniversity of GeorgiaAthensUSA

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