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Fluorescent Real-Time Quantitative Measurements of Intracellular Peroxynitrite Generation and Inhibition
by Jinfang Liao, Jixiang Liu, Ruogu Peng, Yunting Xi, Zhen Luo, Zhenjun Diwu, Jixiang Liu, Qin Zhao
Abstract
Peroxynitrite (ONOO-), a strong oxidant species, is produced by the reaction of nitric oxide (NO) and superoxide (O2•-) radicals. It plays an important role as a biological regulator in numbers of physiological and pathological processes. In this study, we developed fluorescence-based real-time quantitative measurements to detect intracellular ONOO-. The probe DAX-J2 PON Green showed high selectivity toward ONOO- over other competing species, and has been successfully applied in microplate reader and flow cytometer to quantitatively measure endogenous ONOO- production. Moreover, the results demonstrated the inhibitory effects of curcumin on intracellular ONOO- generation.
Keywords: Curcumin; Flow cytometer; Fluorescence; Microplate reader; Peroxynitrite.
Graphical Abstract
Fig. 1
DAX-J2 PON Green labeled RAW 264.7 macrophage cells were treated with SIN-1 to generate intracellular peroxynitrite (ONOO-)
Copyright © 2017 Elsevier Inc. All rights reserved.
https://doi.org/10.1016/j.ab.2017.01.001
Document: 02.0160.200708r1
Last updated Mon Oct 13 2025
Fluorescent Real-Time Quantitative Measurements of Intracellular Peroxynitrite Generation and Inhibition