Intensity-independent fluorometric detection of cellular nitric oxide release

被引：14

作者：

Andrew, PJ

Auer, M

Lindley, IJD

Kauffmann, HF

Kungl, AJ

机构：

[1] NOVARTIS FORSCHUNGSINST,DEPT IMMUNODERMATOL,A-1235 VIENNA,AUSTRIA

[2] UNIV VIENNA,INST PHYS CHEM,A-1090 VIENNA,AUSTRIA

来源：

FEBS LETTERS | 1997年 / 408卷 / 03期

基金：

奥地利科学基金会;

关键词：

nitric oxide; fluorescence spectroscopy; diaminonaphthalene; single-photon counting;

D O I：

10.1016/S0014-5793(97)00449-3

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A new fluorescence method is introduced in which nitric oxide (NO)-derived higher-order oxygen complexes (NOx) are quantified at physiological pH. Detecting the fluorescence lifetime shift between 2,3-diaminonaphthalene and the NOx-derived protonated 2,3-naphthotriazole allows an intensity independent determination of the NOx concentration, The NO release from LPS and IFN gamma-stimulated murine macrophages and iNOS transfected hamster cells was quantified. The lower detection limit for NO2- was found to be 800 pmol/ml, Since the influence of static fluorescence quenching due to cellular components can be neglected, the method is applicable for clear cellular supernatants as well as turbid cellular suspensions. (C) 1997 Federation of European Biochemical Societies.

引用

页码：319 / 323

页数：5

共 17 条

[1]

Berlman I.B., 1971, Handbook of Fluorescence Spectra of Aromatic Molecules

[2] FLUOROMETRIC-DETERMINATION OF NITRITE [J].