Human il 1β il 1f2 duoset kit

Cytokine production by cells in culture was quantified from cell supernatants using either the human or mouse T_H1/T_H2/T_H17 Cytometric Bead Array (BD Bioscience) or via intracellular cytokine staining after treated for 4 hours with PMA (50 ng/ml), ionomycin (1 μg/ml) (both Sigma Aldrich), and 1× Golgi Plug (BD Biosciences). Secreted human IL-1β and IL-18 were measured using the Human IL-1β/IL-1F2 DuoSet Kit or the Human IL-18 Platinum ELISA kit (R&D Systems and eBiosciences, respectively) in combination with SIGMAFAST OPD tablets (Sigma Aldrich) as substrate for detection.

Secreted human IL-1β and IL-10 were measured using the Human IL-1β/IL-1F2 DuoSet Kit or the Human IL-10 Quantikine Elisa kit (R&D Systems, DY201 and D1000B, respectively). Additional cytokines, growth factors and proteins (TNF, G-CSF, IL-6, IL-5, FGF, Arginase, IL-1RA, etc.) were quantified using a custom-assembled Human Multiplex Cytokine Assay (Bio-Plex; Bio-Rad Laboratories Inc.). Mouse IL-1β measured by either utilizing the Mouse IL-1 beta/IL-1F2 DuoSet ELISA kit (R&D Systems, DY401; recognizes predominantly mature IL-1β), via intracellular staining, or with the LEGENDplex^™ Mouse Inflammation Panel (13-plex) (BioLegend, 140150). The latter kit was also used to assess the additional mouse cytokines measured in this study.