Ready set go enzyme linked immunosorbent assay elisa kits

Cytokine panel in the mouse serum were detected with Milliplex Map kit (Millipore, #MCYTOMAG-70K). All assays were performed according to the manufacturer’s protocols, and the MFIs were detected by the Luminex 200 system and were analyzed with Bio-Plex software (Bio-Rad). TNF-α, IL-6, IL-1β levels in the supernatant of the cells were measured with Ready-SET-GO enzyme-linked immunosorbent assay (ELISA) kits (eBioscience). All samples were measured in triplicate according to the manufacturer’s protocol.

Murine TNF-α or Murine IL-10 Ready set go enzyme-linked immunosorbent assay (ELISA) kits were used (eBioscience). Protocol was performed as per manufacturer’s instructions. In brief, Costar 9018 ELISA plates were coated with 100 μL/ well of capture antibody diluted in Coating Buffer overnight at 4°C followed by blocking with ELISASPOT diluent. Small intestinal samples were homogenized in Cell Lytic MT buffer (Sigma) using Lysing matrix E beads (MP Biomedicals) and homogenates cleared by centrifugation at 10 000 rpm for 15 minutes. After quantification using BCA kit (Pierce), equivalent amounts of test samples or standard material were applied to wells in duplicate and incubated overnight at 4°C. Detection Antibody, Streptavidin-HRP and substrate were then applied, and plates were read at 450 nm with 570 nm correction.