过氧化氢
细胞内
化学
活性氧
生物物理学
NADPH氧化酶
荧光
细胞生物学
绿色荧光蛋白
生物分子
氧化应激
生物化学
超氧化物
生物
酶
基因
物理
量子力学
作者
Yulia G. Ermakova,Natalia M. Mishina,Carsten Schultz,Vsevolod V. Belousov
出处
期刊:Methods in molecular biology
日期:2019-01-01
卷期号:: 259-274
被引量:8
标识
DOI:10.1007/978-1-4939-9424-3_15
摘要
Reactive oxygen species (ROS) are involved in regulating normal physiological cell functions as second messengers as well as nonspecific damage of biomolecules in a pathological process known as oxidative stress. The HyPer family of genetically encoded probes are a useful noninvasive tool for monitoring the real-time dynamics of ROS in individual cells or model organisms. HyPer, the first genetically encoded probe for detection of hydrogen peroxide (H2O2), is oxidized with high specificity and sensitivity by H2O2, leading to ratiometric changes in the fluorescence excitation spectrum of the probe. These changes can be detected with a wide range of commercial confocal and wide-field microscope systems. Here we describe a detailed protocol for ratiometric monitoring of H2O2 produced by D-amino acid oxidase (DAAO) or by NADPH oxidase (NOX) in NIH-3T3 cells using the HyPer probe.
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