Quantifying Nitric Oxide in Cell Cultures

The NO₂⁻ concentration in culture media was measured to assess NO production in MGCs using Griess reagent. For each sample, 50 μl aliquots were mixed with 50 μl of the Griess reagent (1% sulfanilamide/0.1% naphthylethylene diamine dihydrochloride/2% phosphoric acid) in a 96-well plate. The absorbance at 550 nm was then measured on a microplate reader (SpectraMax M5; Molecular Devices). NaNO₂ was used as the standard to calculate NO₂ concentrations.

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Kim J.H., Han J., Afridi R., Kim J.H., Rahman M.H., Park D.H., Lee W.S., Song G.J, & Suk K. (2023). A multiplexed siRNA screen identifies key kinase signaling networks of brain glia. Life Science Alliance, 6(5), e202201605.

Publication 2023

SpectraMax M5

Culture media Devices Diamine Griess reagent Phosphoric acid Sulfanilamide

Corresponding Organization : Kyungpook National University

Other organizations : Johns Hopkins University, Johns Hopkins Medicine, Kyungpook National University Hospital, International St. Mary's Hospital, Catholic Kwandong University

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Variable analysis

independent variables

Concentration of NO2- in culture media

dependent variables

Nitric oxide (NO) production in MGCs

control variables

Volume of samples (50 μl aliquots)
Volume of Griess reagent (50 μl)
Composition of Griess reagent (1% sulfanilamide, 0.1% naphthylethylene diamine dihydrochloride, 2% phosphoric acid)
Absorbance measurement at 550 nm using a microplate reader (SpectraMax M5; Molecular Devices)

controls

NaNO2 standard used to calculate NO2 concentrations
Not explicitly mentioned

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