Elisa kit for il 6

MH7A cells (2 × 10⁴) that were transfected with an affibody-encoding plasmid were plated in a 24-well plate and cultured in the presence or absence of 100 ng/mL TNF-α. After 24 or 48 h of incubation, IL-6 and PGE2 levels were determined by assaying supernatants with an Enzyme-Linked Immunosorbent Assay (ELISA) Kit for IL-6 (R&D Systems, Minneapolis, MN, USA) or PGE2 (ENZO Life Sciences, Farmingdale, NY, USA), respectively, according to the instructions.

Ang III and Ang II were purchased from Bachem (Torrance, CA, USA). Tissue culture supplies, such as fetal bovine serum (FBS), Dulbecco’s modified Eagles Medium (DMEM)/F12 (1:1), streptomycin, penicillin and trypsin/ethylenediamine tetraacetic acid (EDTA) were purchased from Fisher Scientific (Milford, MA, USA) or VWR (Suwannee, GA, USA). The AT₁R blocker (Losartan) was provided by Du Pont Merck (Wilmington, DE, USA). The AT₂R blocker (PD123319), and the JAK2 inhibitor (AG490) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Protein measurement supplies, acrylamide, enhanced chemiluminescence (ECL) reagents, nitrocellulose membrane, gel electrophoresis and Western blotting supplies inclusive of bicinchoninic acid (BCA) protein reagents were purchased from either Bio-Rad Laboratories (Hercules, CA, USA) or VWR (Piscataway, NJ, USA). The non-phosphorylated and phospho-specific STAT3 antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). Applied Biosystems (Foster City, CA, USA) supplied reverse transcription reagent kit and TaqMan gene expression primers for rat IL-6. An ELISA kit for IL-6 was purchased from R&D Systems (Minneapolis, MN, USA). All the other chemicals were obtained from either Sigma-Aldrich (St. Louis, MO, USA) or Fisher Scientific (Milford MA, USA) or VWR international (Suwannee, GA, USA).