Human il 8 cxcl8 quantikine elisa kit d8000c

Cytokine and chemokine concentrations in cell culture medium were measured using the Human IL‐8/CXCL8 Quantikine ELISA kit (D8000C) or the Human TNF‐alpha Quantikine HS ELISA kit (HSTA00E) from R&D Systems, Minneapolis, MN, according to the manufacturer's instructions. For IL‐8 measurements, equal numbers of primary fibroblasts from the OTULIN^G281R patient or healthy control were seeded in flat‐bottom 96‐well cell culture plates 24 h before the experiments. Cells were washed, and fresh culture medium was added immediately before the cells were treated with 10 ng/ml recombinant human TNF (Life Technologies) as indicated. The cell culture medium was collected at the indicated times (for the “0” samples, medium from untreated cells was left on the cells for 24 h and collected at the same time at the 24 h TNF‐treated samples) and subjected to ELISA analysis. For TNF measurements, equal numbers of shControl and shOTULIN THP‐1 cells were washed in PBS and seeded in fresh medium in round‐bottom 96‐well plates. The cells were then left untreated, and the culture medium was collected at the indicated times and subjected to ELISA analysis.

Porcine pepsin (EC 3.4.23.1, from porcine gastric mucosa, 3850 U mg⁻¹ protein), pancreatin from porcine pancreas (4 × USP specifications (5.46 U mg⁻¹ based on trypsin activity), EC 232-468-9), Dipeptidyl Peptidase IV (DPP-IV from porcine kidney, EC 3.4.14.5, ≥10 U mg⁻¹ protein), Gly-Pro-p-nitroanilide hydrochloride, Angiotensin Converting Enzyme (ACE from rabbit lung, EC 3.4.15.1), and all other reagents were purchased from Sigma-Aldrich (Sigma-Aldrich, Steinheim, Germany). The Active Glucagon-Like Peptide RIA kit (Cat.# GLP1A-35HK) was purchased from Merck Millipore (Merck-Millipore, Darmstadt, Germany). The Gastrin/CCK kit was purchased from CisBio (CisBio, Saclay, France). Human IL-8/CXCL8 Quantikine ELISA Kit (D8000C) was purchased from R&D Systems (R&D Sytems, Minneapolis, MN, USA).