Intensity-independent fluorometric detection of cellular nitric oxide release

P.J. Andrew; M. Auer; I.J.D. Lindley; A.J. Kungl; H.F. Kauffmann

doi:10.1016/S0014-5793(97)00449-3

Intensity-independent fluorometric detection of cellular nitric oxide release

P.J. Andrew, M. Auer, I.J.D. Lindley, A.J. Kungl, H.F. Kauffmann

School of Biological Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

A new fluorescence method is introduced in which nitric oxide (NO)-derived higher-order oxygen complexes (NO(x)) are quantified at physiological pH. Detecting the fluorescence lifetime shift between 2,3-diaminonaphthalene and the NO(x)-derived protonated 2,3-naphthotriazole allows an intensity independent determination of the NO(x) concentration. The NO release from LPS and IFNγ-stimulated murine macrophages and iNOS transfected hamster cells was quantified. The lower detection limit for NO 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.

Original language	English
Pages (from-to)	319-323
Number of pages	5
Journal	FEBS Letters
Volume	408
Issue number	3
DOIs	https://doi.org/10.1016/S0014-5793(97)00449-3
Publication status	Published - 26 May 1997

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title = "Intensity-independent fluorometric detection of cellular nitric oxide release",

abstract = "A new fluorescence method is introduced in which nitric oxide (NO)-derived higher-order oxygen complexes (NO(x)) are quantified at physiological pH. Detecting the fluorescence lifetime shift between 2,3-diaminonaphthalene and the NO(x)-derived protonated 2,3-naphthotriazole allows an intensity independent determination of the NO(x) concentration. The NO release from LPS and IFNγ-stimulated murine macrophages and iNOS transfected hamster cells was quantified. The lower detection limit for NO 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.",

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AU - Auer, M.

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AU - Kungl, A.J.

AU - Kauffmann, H.F.

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N2 - A new fluorescence method is introduced in which nitric oxide (NO)-derived higher-order oxygen complexes (NO(x)) are quantified at physiological pH. Detecting the fluorescence lifetime shift between 2,3-diaminonaphthalene and the NO(x)-derived protonated 2,3-naphthotriazole allows an intensity independent determination of the NO(x) concentration. The NO release from LPS and IFNγ-stimulated murine macrophages and iNOS transfected hamster cells was quantified. The lower detection limit for NO 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.

AB - A new fluorescence method is introduced in which nitric oxide (NO)-derived higher-order oxygen complexes (NO(x)) are quantified at physiological pH. Detecting the fluorescence lifetime shift between 2,3-diaminonaphthalene and the NO(x)-derived protonated 2,3-naphthotriazole allows an intensity independent determination of the NO(x) concentration. The NO release from LPS and IFNγ-stimulated murine macrophages and iNOS transfected hamster cells was quantified. The lower detection limit for NO 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.

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Intensity-independent fluorometric detection of cellular nitric oxide release

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