Human proinflammatory 9 plex ultrasensitive kit

Blood samples were sent for immediate analysis of plasma glucose, serum insulin and insulin-like growth factor 1 (IGF-1). Serum was frozen (−70°C) to allow for analysis of each participant’s blood lipids (total cholesterol, triglycerides, HDL-cholesterol, LDL-cholesterol) and high sensitive C-reactive protein (hs-CRP) from before and after intervention in the same batch to reduce measurement errors. These analyses were performed at the Central Laboratory at the Sahlgrenska University Hospital. Two markers of inflammation (IL-6 and TNF-α) were measured in serum with the Human Proinflammatory 9-plex Ultrasensitive Kit in a SECTOR 2400 Imager (Meso Scale Discovery, Gaithersburg, MD) at the Wallenberg Laboratory for Cardiovascular and Metabolic Research, University of Gothenburg, Sweden.

A multiplex immunoassay was used to measure the levels of nine cytokines in the incubation medium: granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon-γ (IFN-γ), IL-1β, IL-2, IL-6, IL-8, IL-10, IL-12p70, and TNF-α (Human Pro-Inflammatory 9-Plex Ultra-Sensitive Kit, Meso Scale Discovery, Gaithersburg, MD). ELISA was used to measure adiponectin (Human adiponectin ELISA kit, Millipore, Billerica, MA), chemerin (Human chemerin ELISA kit, Millipore), omentin (Human omentin-1 ELISA, Millipore), IL-6 (Quantikine High Sensitivity Human IL-6 ELISA, R&D Systems, Minneapolis, MN), and DPP4 (Quantikine Human DPPIV/CD26 ELISA kit, R&D Systems). Adipokine concentrations were expressed per g adipose tissue.