ArticleNitric oxide : biology and chemistry2013
Kinetic analysis of DAF-FM activation by NO: toward calibration of a NO-sensitive fluorescent dye.
Article in Nitric oxide : biology and chemistry, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 60 citations in OpenAlex.
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- PLLA Composites Combined with Delivery System of Bioactive Agents for Anti-Inflammation and Re-Endothelialization.Pharmaceutics · 2022Article
- Interactions between Nitric Oxide and Hyaluronan Implicate the Migration of Breast Cancer Cells.Biomacromolecules · 2022Article
- Low-Dose Tacrolimus Promotes the Migration and Invasion and Nitric Oxide Production in the Human-Derived First Trimester Extravillous Trophoblast Cells In Vitro.International journal of molecular sciences · 2022Article
- High-throughput production of functional prototissues capable of producing NO for vasodilation.Nature communications · 2022Article
- Hexokinase 2 Inhibition and Biological Effects of BNBZ and Its Derivatives: The Influence of the Number and Arrangement of Hydroxyl Groups.International journal of molecular sciences · 2022Article
- New Laboratory Protocol to Determine the Oxidative Stress Profile of Human Nasal Epithelial Cells Using Flow Cytometry.Journal of clinical medicine · 2021Article
- Antimicrobial carbon nanodots: photodynamic inactivation and dark antimicrobial effects on bacteria by brominated carbon nanodots.Nanoscale · 2021Article
- A DNA-based fluorescent probe maps NOS3 activity with subcellular spatial resolution.Nature chemical biology · 2020Article
- Human In Vitro Models for Assessing the Genomic Basis of Chemotherapy-Induced Cardiovascular Toxicity.Journal of cardiovascular translational research · 2020Review
- Incubation with sodium nitrite attenuates fatigue development in intact single mouse fibres at physiologicalThe Journal of physiology · 2019Article
- Light-induced release of nitric oxide from the nitric oxide-bound CDGSH-type [2Fe-2S] clusters in mitochondrial protein Miner2.Nitric oxide : biology and chemistry · 2019Article
- Role of nitric oxide in pancreatic cancer cells exhibiting the invasive phenotype.Redox biology · 2019Article
- Edible Bird's Nest Protects Against Hyperglycemia-Induced Oxidative Stress and Endothelial Dysfunction.Frontiers in pharmacology · 2019Article
- Arginine and the metabolic regulation of nitric oxide synthesis in cancer.Disease models & mechanisms · 2018Review
- Regulation of Cellular Redox Signaling by Matricellular Proteins in Vascular Biology, Immunology, and Cancer.Antioxidants & redox signaling · 2017Article
- Granular Layer Neurons Control Cerebellar Neurovascular Coupling Through an NMDA Receptor/NO-Dependent System.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2017Article
- Development of High-Throughput Method for Measurement of Vascular Nitric Oxide Generation in Microplate Reader.Molecules (Basel, Switzerland) · 2017Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Nitric oxide (NO) research in biomedicine has been hampered by the absence of a method that will allow quantitative measurement of NO in biological tissues with high sensitivity and selectivity, and with adequate spatial and temporal resolution. 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM) is a NO sensitive fluorescence probe that has been used widely for qualitative assessment of cellular NO production. However, calibration of the fluorescent signal and quantification of NO concentration in cells and tissues using fluorescent probes, have provided significant challenge. In this study we utilize a combination of mathematical modeling and experimentation to elucidate the kinetics of NO/DAF-FM reaction in solution. Modeling and experiments suggest that the slope of fluorescent intensity (FI) can be related to NO concentration according to the equation: ddtFI=2αk(1)NO(2)O(2)DAF-FMkNO+DAF-FM where α is a proportionality coefficient that relates FI to unit concentration of activated DAF-FM, k(1) is the NO oxidation rate constant, and k was estimated to be 4.3±0.6. The FI slope exhibits saturation kinetics with DAF-FM concentration. Interestingly, the effective half-maximum constant (EC(50)) increases proportionally to NO concentration. This result is not in agreement with the proposition that N(2)O(3) is the NO oxidation byproduct that activates DAF-FM. Kinetic analysis suggests that the reactive intermediate should exhibit NO-dependent consumption and thus NO(2)() is a more likely candidate. The derived rate law can be used for the calibration of DAF-FM fluorescence and the quantification of NO concentration in biological tissues.
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