Enzyme linked immunosorbent assay elisa kits for pge2

DMC (Figure 1A(Fig. 1)), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Invitrogen-Gibco (Grand Island, NY, USA). Enzyme-linked immunosorbent assay (ELISA) kits for PGE₂, TNF-α, IL-1ß, and IL-6 were purchased from R&D Systems, Inc. (St. Louis, MO, USA). Anti-iNOS (mouse), Anti-COX-2 (rabbit), Anti-IκB-α (mouse), anti-phosphorylated IκB-α (rabbit, anti-p-IκB-α), anti-c-Jun N-terminal kinase (rabbit, JNK), anti-phosphorylated JNK (rabbit, anti-p-JNK), anti-extracellular signal-regulated kinases (rabbit, ERK1/2), anti-phosphorylated ERK1/2 (rabbit, anti-p-ERK1/2), anti-p38 (rabbit), and anti-phosphorylated p38 (rabbit, anti-p-p38) mouse or rabbit antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). All other reagents were purchased from the Sigma-Aldrich Chemical Co. (St. Louis, MO, USA).

Selagin-7-O-(6"-O-Acetyl-)-ß-D-glucoside (Figure 1(Fig. 1)) used for this study was isolated from C. discoidea (Zhu et al., 2010[35 ]). The isolated compound was analyzed by liquid chromatography coupled with mass spectrometry, and its purity was found to be > 99 %. Dimethyl sulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-y1)-2, 5-diphenyltetrazolium bromide (MTT), and PBS were purchased from Sigma–Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were obtained from Invitrogen-Gibco (Grand Island, NY, USA). Enzyme-linked immunosorbent assay (ELISA) kits for PGE2, TNF-α, IL-1ß, and IL-6 were purchased from R&D Systems, Inc. (St. Louis, MO, USA), and ELISA kits for interleukin-10 (IL-10) was purchased from Biolegend (CA, USA). The antibody to NF-κB p65 was purchased from Biovision (Mountain View, CA, USA). The antibodies to COX-2 and iNOS were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and the antibody to ß-actin was purchased from Chemicon (Temecula, CA, USA). The other chemicals and reagents used were of analytical grade.