Application of 4,5-diaminofluorescein to reliably measure nitric oxide released from endothelial cellsin vitroReport as inadecuate




Application of 4,5-diaminofluorescein to reliably measure nitric oxide released from endothelial cellsin vitro - Download this document for free, or read online. Document in PDF available to download.

Biological Procedures Online

, Volume 5, Issue 1, pp 136–142

Received: 25 February 2003Revised: 31 March 2003Accepted: 19 May 2003DOI: 10.1251-bpo55

Cite this article as: Räthel, T.R., Leikert, J.F., Vollmar, A.M. et al. Biol. Proced. Online 2003 5: 136. doi:10.1251-bpo55

Abstract

Here we describe in more depth the previously published application of the fluorescent probe 4,5-diaminofluorescein DAF-2 in order to reliably measure low levels of nitric oxide NO as released from human endothelial cells invitro. The used approach is based on the following considerations a use low concentrations of DAF-2 0.1 µM in order to reduce the contribution of DAF-2 auto-fluorescence to the measured total fluorescence, and b subtract the DAF-2 auto-fluorescence from the measured total fluorescence. The advantage of this method is the reliable quantification of NO in a biological system in the nanomolar range once thoroughly validated. Here we focus in addition to the previous publication Leikertet al.,FEBS Lett 2001, 506:131–134 on aspects of validation procedures as well as limitations and pitfalls of this method.

Indexing termsInvestigative Techniques Biological Assay Nitric-Oxide Synthase Published: June 2, 2003

Download to read the full article text



Author: Thomas R. Räthel - Jürgen F. Leikert - Angelika M. Vollmar - Verena M. Dirsch

Source: https://link.springer.com/







Related documents