3.8. Nitric Oxide Production Measurements in Macrophage Cells

The amount of stable nitrite, which is the final nitric oxide product generated in LPS-stimulated macrophages, was determined with a colorimetric assay after 24 h of cell seeding (1 × 10⁵ cells/well in 96-well plate). Briefly, 100 μL of the culture supernatant was mixed with an equal volume of Griess reagent (1% sulfanilamide, 0.1% naphthylethylenediamine dihydrochloride, 2.5% H₃PO₄) and incubated at room temperature for 10 min. The absorbance at 550 nm was read using a spectrophotometer.