ArticleBiochemical and biophysical research communications2015
Detection of nitric oxide production in cell cultures by luciferin-luciferase chemiluminescence.
Article in Biochemical and biophysical research communications, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed, 26 citations in OpenAlex.
- A flexible and highly sensitive organic electrochemical transistor-based biosensor for continuous and wireless nitric oxide detection.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Quantitative determination of nitric oxide from tissue samples using liquid chromatography-Mass spectrometry.MethodsX · 2021Article
- A flexible and physically transient electrochemical sensor for real-time wireless nitric oxide monitoring.Nature communications · 2020Article
- Hydrodynamic shear stress promotes epithelial-mesenchymal transition by downregulating ERK and GSK3β activities.Breast cancer research : BCR · 2019Article
- N-acetyl ornithine deacetylase is a moonlighting protein and is involved in the adaptation of Entamoeba histolytica to nitrosative stress.Scientific reports · 2016Article
- Nitric oxide detection methodsMedical gas researchReview
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
Abstract
A chemiluminescent method is proposed for quantitation of NO generation in cell cultures. The method is based on activation of soluble guanylyl cyclase by NO. The product of the guanylyl cyclase reaction, pyrophosphate, is converted to ATP by ATP sulfurylase and ATP is detected in a luciferin-luciferase system. The method has been applied to the measurement of NO generated by activated murine macrophages (RAW 264.7) and bovine aortic endothelial cells. For macrophages activated by lipopolysaccharide and γ-interferon, the rate of NO production is about 100 amol/(cell·min). The rate was confirmed by the measurements of nitrite, the product of NO oxidation. For endothelial cells, the basal rate of NO generation is 5 amol/(cell·min); the rate approximately doubles upon activation by bradykinin, Ca(2+) ionophore A23187 or mechanical stress. For both types of cells the measured rate of NO generation is strongly affected by inhibitors of NO synthase. The sensitivity of the method is about 50 pM/min, allowing the registration of NO generated by 10(2)-10(4) cells. The enzyme-linked chemiluminescent method is two orders of magnitude more sensitive than fluorescent detection using 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM).
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Registered trials
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