Human il 37 elisa kit

H1299, H1299-Mock, and two clones of IL-37-transfected H1299 cells with the same beginning numbers were cultured with same volume of medium and the growth medium supernatant were collected when cells reached 90 % confluence. The concentrations of secreted IL-37 and VEGF in the growth medium supernatants were quantified by using a commercial human IL-37 ELISA kit (AdipoGen AG, Liestal, Switzerland) and VEGF Kit (R&D Systems, Inc., Minneapolis, MN, USA) according to the manufacturer's protocol. All samples were assayed in duplicate.

The level of circulating IL-37 in patients and mice was determined using a human IL-37 ELISA kit (Adipogen, Switzerland). Measurements were performed in duplicate and the results were averaged. Samples with obvious hemolysis, which was visually detected as a pink to red tinge, were not used for measurements. 100 µl of neat plasma was placed into wells coated with anti-human IL-37 antibody in duplicate. Plates were then sealed, incubated overnight at 4 °C and washed, followed by addition of Detection Antibody (Adipogen). Plates were washed again and incubated at 37 °C for 1 h. Horse radish peroxidase-conjugated anti-rabbit IgG was added, the plate was washed again and 3,3′,5,5′-tetramethylbenzidine substrate solution (Adipogen) was added to each well for 10 min in the dark to allow for colour development. Stop solution was then added and absorption was read at 450 nm by a CLARIOstar microplate reader (BMG Labtech, Germany). The standard curve was interpolated in a quadratic equation.