Il 6 elisa kit

The levels of IL-6 and IL-1β in growth medium exposed to AEC culture for 1, 2, and 3 days were analysed by enzyme-linked immunosorbent assay (ELISA) according to the manufacturer’s instructions. IL-6 ELISA kit was purchased from RayBiotech (Georgia, US), while IL-1β ELISA kit was purchased from R&D System (Minneapolis, US). The medium collected was proportioned into 200 µl aliquots and stored at −80 °C. 100 µl of standards and samples (3000 pg/ml) were used for the assay procedure.

Blood samples were collected on the same day of the medical visit. Patients were instructed to fast overnight and not to take any medications before blood sampling on the morning of the tests. Blood samples were drawn after a 30 min supine rest. The following analyses were performed: high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), NT-proBNP, Hb1Ac, and uric acid. Hs-CRP was measured with a quantitative turbidimetric test (TRUEchemie). NT-proBNP was measured with the ECLIA monoclonal assay using the Cobas e411 platform (Roche Diagnostics Italia, Monza, Italy). Plasma norepinephrine was evaluated using high-performance liquid chromatography with the electrochemical detector CLC 100 (Chromsystems, Munchen, Germany). Interleukin-6 was measured with an in vitro enzyme-linked immunosorbent assay for quantitative measurement (RayBiotech IL-6 ELISA kit).