Legend max human il 33 elisa kit

IL-33 plasma concentrations were evaluated using ELISA kit according to the manufacturer’s instruction (LEGEND MAX™ Human IL-33 ELISA Kit, Biolegend).

Serum IL-33 and sST2 concentrations were determined in duplicate using ELISA kits to detect the C-terminal epitope of IL-33 (LEGEND MAX™ Human IL-33 ELISA Kit, BioLegend; DuoSet IL-33 ELISA kit, R&D Systems, Minneapolis, MN, USA) and sST2 (R&D Systems), respectively, in accordance with the manufacturer’s recommended protocols. Briefly, serum samples were incubated in 96-well plates that had previously been coated with anti-IL-33 and sST2-specific monoclonal antibodies for 2 hours at room temperature. After washing the wells four times, peroxidase-conjugated polyclonal antibodies against IL-33 and sST2 were added. Samples were incubated for 2 hours at room temperature followed by four washes, after which the substrate solution was added. After incubation for 30 minutes at room temperature, the reactions were terminated by the addition of stop solution. The optical density of each well at 450 nm was determined using a VersaMax ELISA Microplate Reader (Molecular Devices, Sunnyvale, CA, USA). All values were corrected by normalising to the corresponding absorbances at 540 nm. All data were analysed with SoftMax Pro 6.3 (Molecular Devices).