4.5. Nitric Oxide (NO) Production in Raw 264.7 Cells

The production of NO in LPS-induced cells was measured as an indicator of NO production based on the Griess reaction. The cell culture plate conducted centrifugation process at 2500× g, 5 min. Then, 50 µL of the cell culture supernatant was mixed with 50 µL 1% sulfanilamide, and 0.1% NED incubated at 24 °C for 10 min. The absorbance at 540 nm was measured in an ELISA reader (multiscan spectrum). The amount of nitrite in the samples was determined using sodium nitrite serial dilution standard curve, and nitrite production was measured.