Endo free purification kit

LPA (1-oleoly, 18:1) was obtained in chloroform solvent from Avanti Polar Lipids, Inc. (Alabaster, AL). Prior to use, LPA was dried and dissolved in PBS containing 0.5% fatty acid-free bovine serum albumin (BSA) from Roche (Indianapolis, IN). The ELISA Max™ Deluxe Sets for TNF-α, IL-1β, IL-6 and IL-8 were purchased from BioLegend (San Diego, CA). The corresponding ELISA kits were prepared according to the manufacturer. The human CXCL1 DuoSet ELISA kit was from R&D Systems, Inc. (Minneapolis, MN). The TaqMan Universal PCR Master Mix and qPCR probes for TNF-α, IL-1β and GAPDH were obtained from ThermoFisher Scientific (Waltham, MA). The ADAM17 inhibitor TMI-1, NF-κB inhibitor (BAY11–7085), EGFR inhibitor AG1478, and anti-ADAM17 antibody (ABT94) were obtained from Sigma-Aldrich (St. Louis, MO). Pertussis toxin (PTX) was purchased from List biological labs (Campbell, CA). Anti-EGFR antibody and anti-phospho-EGFR (Y1068) antibody were from Cell Signaling Technology (Danvers, MA). Anti-IL6 antibody (SC-7920) and COX-2 antibody (SC-1745) were from Santa Cruz (Austin, TX). Anti-TNF-α neutralizing monoclonal antibody (TNF-α Ab) was from BioLegend. The soluble form of the TNF-α receptor (TNF-α SR) was obtained from Amgen (Thousand Oaks, CA). Plasmid DNAs were purified using the endo-free purification kit from Qiagen (Valencia, CA).