Pge2 elisa assay kit

Lipopolysaccharide (LPS from Salmonella enterica serotype Typhimurium), indomethacin, and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle Medium (DMEM), 0.25% trypsin-EDTA solution, penicillin/streptomycin (P/S) solution, and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). Dimethyl sulfoxide (DMSO) was purchased from Merck (Darmstadt, Germany). PGE₂ ELISA assay kit was obtained from R&D Systems (Minneapolis, MN, USA). TNF-α ELISA assay kit (ab100747) and nitric oxide assay kit (ab65328) were purchased from Abcam (Waltham, MA, USA). The specific primers were obtained from Integrated DNA Technologies (Coralville, IA, USA). KAPA SYBR FAST One-step RT-qPCR kit was purchased from KAPA biosystems (Wilmington, MA, USA). RNeasy kit was obtained from Qiagen (Hilden, Germany).

Cells were placed in 6-well plates at a density of 80%-90% and subsequently the supernatant was collected after 24 h of incubation in serum-free DMEM. Cellular debris was removed via centrifugation before samples were frozen at -80°C. A PGE2 ELISA Assay kit (JLC5387; R&D Systems, Inc., Minneapolis, MN, USA) was then used to quantify the amount of prostaglandin E2 (PGE2) in the substance, based on protocols stipulated by the manufacturers.

The levels of PGE₂ in the conditioned media were determined by using the PGE₂ ELISA assay kit (R & D Systems) according to the manufacturer’s instructions [16 (link)].