Elisa kits for human il 1β

Penicillin-streptomycin antibiotic, FBS, PBS, and RPMI-1640 culture medium were attained from Gibco (Grand Island, NY, USA). R & D Systems (Minneapolis, MN, USA) were the supplier of ELISA kits for human IL-1β, TNF-α and PGE₂. MTT reagent, LPS, RIPA, and DMSO were obtained from Sigma Chemical Co. (St. Louis, MO, USA). Levamisole (purity > 98%) was procured from Cayman Chemical (Ann Arbor, MI, USA). Tocris Biosciences (Bristol, UK) supplied Akt inhibitor (LY294002), p38 inhibitor (SB202190), ERK inhibitor (U0126), JNK inhibitor (SP600125), and NF-κB inhibitor (BAY 11–7082). Pierce (Rockford, IL, USA) supplied 1 × Halt phosphatase and protease inhibitor cocktail. Cell Signaling Technology (Beverly, MA) supplied primary antibodies specific to p-NFκBp65, p-IKKα/β, p-JNK, IκBα, p-IκBα, p-ERK, p-p38, JNK, ERK, p38, COX-2, p-Akt, TRL4 and MyD88 along with anti-rabbit secondary antibody conjugated to horseradish peroxidase and β-actin.

To quantitate secreted cytokines, cell culture supernatants of BMDMs or peritoneal lavage fluids were measured by ELISA Kits for human IL-1β (DY201-05, R&D Systems), mouse IL-1β (DY401, R&D Systems), mouse IL-6 (M6000B, R&D Systems), and mouse IL-18 (BMS618/3, eBioscience, San Diego, CA, USA). The ELISA plates were readout using a Synergy™ H1 Hybrid Multi-Mode Reader, BioTek).