Document Detail

Reliable in vitro measurement of nitric oxide released from endothelial cells using low concentrations of the fluorescent probe 4,5-diaminofluorescein.
MedLine Citation:
PMID:  11591386     Owner:  NLM     Status:  MEDLINE    
4,5-Diaminofluorescein (DAF-2) and its membrane-permeable derivate DAF-2 diacetate are fluorescent probes that have been developed to perform real-time biological detection of nitric oxide (NO). Their use for intracellular imaging, however, has recently been seriously questioned and data using DAF-2 for extracellular NO detection at low levels, as for example released from endothelial cells, are rare. Here we show that a reliable detection of low levels of NO in biological systems by DAF-2 is possible (a) by using low DAF-2 concentrations (0.1 microM) and (b) by subtracting the DAF-2 auto-fluorescence from the measured total fluorescence. The described method allows easy real-time detection of endothelial NO formation.
J F Leikert; T R Räthel; C Müller; A M Vollmar; V M Dirsch
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  FEBS letters     Volume:  506     ISSN:  0014-5793     ISO Abbreviation:  FEBS Lett.     Publication Date:  2001 Oct 
Date Detail:
Created Date:  2001-10-09     Completed Date:  2001-12-04     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  0155157     Medline TA:  FEBS Lett     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  131-4     Citation Subset:  IM    
Department of Pharmacy, Center of Drug Research, University of Munich, Butenandtstr. 5-13, D-81377, Munich, Germany.
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MeSH Terms
Cell Line
Endothelium, Vascular / chemistry*,  cytology,  drug effects
Enzyme Inhibitors / pharmacology
Fluorescent Dyes
Molecular Structure
Nitric Oxide / analysis*
Spectrometry, Fluorescence
Tetradecanoylphorbol Acetate / pharmacology
Triazenes / metabolism
omega-N-Methylarginine / pharmacology
Reg. No./Substance:
0/4,5-diaminofluorescein; 0/Enzyme Inhibitors; 0/Fluorescent Dyes; 0/Triazenes; 10102-43-9/Nitric Oxide; 146724-86-9/NOC 9; 16561-29-8/Tetradecanoylphorbol Acetate; 17035-90-4/omega-N-Methylarginine; 2321-07-5/Fluorescein

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