Protocols to Measure Oxidative Stress and DNA Damage in Asthma

Koa Hosoki, Anirban Chakraborty, Tapas K. Hazra, Sanjiv Sur

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Asthma is associated with oxidative stress and oxidative damage of biomolecules, including DNA. Here, we describe the protocols to quantify reactive oxygen species (ROS) and oxidative stress markers in a mouse model of allergic airway inflammation. We also provide detailed methods to measure DNA damage by long-run real-time PCR for DNA-damage quantification (LORD-Q) assay and gene-specific DNA damage analyses by long amplicon (LA)-qPCR. Additionally, we describe methods to quantify oxidized DNA base lesions in lung genomic DNA by mass spectrometry, and to measure enzymatic activity of 8-oxoguanine DNA glycosylase (OGG1). Using these methods, the levels of oxidative stress and DNA damage in allergic inflammation and asthma can be elucidated.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages315-332
Number of pages18
DOIs
StatePublished - Jul 1 2022

Publication series

NameMethods in Molecular Biology
Volume2506
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Allergic inflammation
  • Asthma
  • GSSG
  • LA-PCR
  • LORD-Q PCR
  • NEIL2
  • OGG1
  • Oxidative DNA damage
  • Reactive oxygen species

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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