Pge2 immunoassay kit

Antibodies specific for EP2, EP3, EP4, Sp1, Sp3, TET2, TET3, β-actin, vinculin, and histone H3 were purchased from Santa Cruz Biotechnology (Dallas, TX). The following antibodies were purchased: COX-2 and 5-methylcytosine (5mC) from Cell Signaling (Danvers, MA); EP1 from Abcam (Cambridge, MA); Dnmt1, Dnmt3a, and Dnmt3b from Novus Biologicals (Littleton, CO); and TET1 from Epigentek, Inc (New York, NY). The Methylamp Global DNA Methylation Quantification Kit, the EpiQuik DNA Methyltransferase Activity Assay Kit, and TET Assay Activity Kit (Epigenase 5mC-hydroxylase TET activity/inhibition assay kit) were purchased from Epigentek, Inc. (New York, NY). The PGE₂ immunoassay kit was purchased from Cayman Chemical (Ann Arbor, MI). All other chemicals of analytical grade were purchased from Sigma-Aldrich Chemical Co (St Louis, MO). Purified honokiol was purchased from Quality Phytochemicals, LLC (Edison, NJ).

MDR-Caco-2 and Caco-2 cells at a density of 5×10⁶ were seeded in 90 mm culture dishes. They were incubated with or without snomenine (500 µM) for 48 h. At the end of the treatment period, culture medium was collected to determine the amount of PGE₂ secreted by these cells and stored at)−80°C. The quantitative analysis of PGE₂ released into the medium was assessed using PGE₂ immunoassay kit as per manufacturer’s instructions (Cayman Chemical Company, USA).