title: Fluorescent real-time quantitative measurements of intracellular peroxynitrite generation and inhibition.
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.
authors.affiliation: AAT Bioquest, Inc., United States. Electronic address: zhenluo@aatbio.com.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Luo
authors.affiliation: AAT Bioquest, Inc., United States.
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: AAT Bioquest, Inc., United States.
authors.forename: Jixiang
authors.initials: J
authors.lastname: Liu
authors.affiliation: AAT Bioquest, Inc., United States.
authors.forename: Jinfang
authors.initials: J
authors.lastname: Liao
authors.affiliation: AAT Bioquest, Inc., United States.
authors.forename: Ruogu
authors.initials: R
authors.lastname: Peng
authors.affiliation: AAT Bioquest, Inc., United States.
authors.forename: Yunting
authors.initials: Y
authors.lastname: Xi
authors.affiliation: AAT Bioquest, Inc., United States.
authors.forename: Zhenjun
authors.initials: Z
authors.lastname: Diwu
mesh_terms: D000818:Animals; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D008856:Microscopy, Fluorescence; D009569:Nitric Oxide / Q000737:chemistry; D030421:Peroxynitrous Acid / Q000032:analysis* / Q000378:metabolism; D000067996:RAW 264.7 Cells; D013481:Superoxides / Q000737:chemistry; D046888:Tissue Array Analysis
chemical_list: D005456:Fluorescent Dyes; D013481:Superoxides; D030421:Peroxynitrous Acid; D009569:Nitric Oxide
keywords: Curcumin; Flow cytometer; Fluorescence; Microplate reader; Peroxynitrite