Journal
FREE RADICAL BIOLOGY AND MEDICINE
Volume 46, Issue 7, Pages 853-865Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.freeradbiomed.2008.12.020
Keywords
Reactive oxygen species; Free radicals; Spin trap; Immuno-spin trapping; Immunoassay; Mass spectrometry; Confocal microscopy; Molecular resonance imaging
Funding
- National institute of Environmental Health Sciences [R00ES015415]
- Intramural Research Program of the NIEHS/NIH
- Presbyterian Health Foundation
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Biomolecule-centered radicals are intermediate species produced during both reversible (redox modulation) and irreversible (oxidative stress) oxidative modification of biomolecules. These oxidative processes must be Studied in situ and in real time to understand the molecular mechanism of cell adaptation or death in response to changes in the extracellular environment. In this regard, we have developed and validated immuno-spin trapping to tag the redox process, tracing the oxidatively generated modification of biomolecules, in situ and in real time, by detecting protein- and DNA-centered radicals. The purpose of this methods article is to introduce and Update the basic methods and applications of immuno-spin trapping for the study of redox biochemistry in oxidative stress and redox regulation. We describe in detail the production, detection, and location of protein and DNA radicals in biochemical systems, cells, and tissues, and in the whole animal as well, by using immuno-spin trapping with the nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide. (C) 2009 Elsevier Inc. All rights reserved.
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