AAT Bioquest

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


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

OxiVision Blue

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.

Original created on July 8, 2020, last updated on July 8, 2020
Tagged under: Intracellular Peroxynitrite Detection