4.5. Measurement of Cytokines by ELISA

The secretion of pro-inflammatory cytokines, including IL-6 and TNF-α, was measured with an ELISA assay kit (R&D Systems, Minneapolis, MN, USA). RAW 264.7 cells were seeded into a 96-well plate containing 3–100 μM SNAH with or without 0.2 μg/mL LPS for 6 h. The supernatant was harvested, and pro-inflammatory cytokine levels were determined using ELISA kits. Absorbance was measured at 450 nm using a microplate spectrophotometer (Bio-Tek). The quantities of cytokines secreted by RAW 264.7 cells were calculated using standard curves.