Pge2 monoclonal enzyme immunoassay eia kit

Following the same treatments as described above, the level of IL8 in the culture supernatants was determined using IL8 human ELISA kit (ThermoFisher Scientific); and the PGE2 level was determined using PGE2 monoclonal enzyme immunoassay (EIA) kit (Cayman Chemicals, Ann Arbor, MI, USA) according to the manufacturer’s instructions.

For measurement of inflammatory mediators, cells (4 × 10⁴) were seeded in 24-well plates in 300 μL medium and incubated for 24 hours to allow attachment. The medium was removed and cells were rinsed twice with serum-free medium and thereafter stimulated in serum-free media with IL-1β (300 pg/mL) alone or in combination with increasing subcytotoxicity concentrations of the oils. Supernatants were collected after 24 hours of incubation and stored at −80°C until analysis. IL-6 and IL-8 levels in the supernatants were measured using the Duoset ELISA kit (R&D Systems Inc., Minneapolis, MN, USA) and PGE₂ levels determined using PGE₂ monoclonal enzyme immunoassay (EIA) kit (Cayman Chemicals, Ann Arbor, MI, USA). Both kits were used according to the manufacturer's instructions. Readings were made at 450 nm for IL-6 and IL-8 and at 405 nm for PGE₂ with microplate spectrophotometer (Labsystem Multiskan MS).