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Measuring Nitric Oxide in Cell Supernatant
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Cell supernatant nitric oxide levels were determined by Griess assay. Cell supernatant or standards were added in equal volumes with Griess Reagent (1:1 ratio of 2% sulphanilamide in 5% H3PO4 and 0.2% Napthylene diamine HCL in ddH2O) in a 96 well-plate and incubated in the dark for 10 min. Absorbance was read at 540 nm on a Spectromax 190 plate reader and serum nitrite concentrations were calculated against a standard curve.
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