A nitric oxide quantitative assay by a glyceraldehyde 3-phosphate dehydrogenase/phosphoglycerate kinase/firefly luciferase optimized coupled bioluminescent assay

Marques, Simone M.; Silva, Joaquim C. G. Esteves da

doi:10.1039/c4ay00317a

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2023

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Cited by 2 publications

(2 citation statements)

References 49 publications

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“…Concentrations after release were calculated by considering a ratio of 1.5 moles of NO per parent compound, according to the manufacturer's information. 32…”

Section: Synthetic Proceduresmentioning

confidence: 99%

Direct fluorescence labelling of NO inside plant cells

Ghosh,

Saha,

Mukherjee

et al. 2023

Org. Biomol. Chem.

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“…Concentrations after release were calculated by considering a ratio of 1.5 moles of NO per parent compound, according to the manufacturer's information. 32…”

Section: Synthetic Proceduresmentioning

confidence: 99%

Direct fluorescence labelling of NO inside plant cells

Ghosh,

Saha,

Mukherjee

et al. 2023

Org. Biomol. Chem.

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“…Enzyme-linked NO detection suggested in our and other works [15–17] is based on the changing of catalytic activity of an enzyme in the presence of NO. The concept, first suggested in our publication [17], is outlined on Scheme 1.…”

Section: Introductionmentioning

confidence: 96%

Detection of nitric oxide production in cell cultures by luciferin–luciferase chemiluminescence

Woldman

Eubank

Mock

et al. 2015

Biochemical and Biophysical Research Communications

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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, Ca2+ 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 102–104 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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A nitric oxide quantitative assay by a glyceraldehyde 3-phosphate dehydrogenase/phosphoglycerate kinase/firefly luciferase optimized coupled bioluminescent assay

Abstract: A fast, simple, sensitive and robust enzymatic method with bioluminescent detection for nitric oxide is presented.

Cited by 2 publications

References 49 publications

Direct fluorescence labelling of NO inside plant cells

Direct fluorescence labelling of NO inside plant cells

Detection of nitric oxide production in cell cultures by luciferin–luciferase chemiluminescence

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