Elisa max standard set human il 6

For the Caco2 cell transwell experiments, f-HMOs were incubated for 24 h and the apical and basolateral media were harvested for cytokine analysis. Cytokine secretion was quantified using Meso Scale Discovery (MSD, Rockville, MD, USA). IL-6, IL-8, and GRO-α were quantified simultaneously on one plate using the MSD U-plex Biomarker Group 1 (hu) assay. TGF-β1, -2, and -3 were quantified simultaneously using the MSD U-PLEX TGF-β Combo (hu) assay. The plates were read using the 1300 Meso QickPlex SQ 120 (MSD, Rockville, MD, USA). The data were expressed as total cytokine amounts secreted (in pg).
For the gut-on-chip experiments, apical and basolateral effluents from Colon Intestine-Chips were analyzed for IL-6 and IL-8 secretion using ELISA MAX Standard Set Human IL-6 (Biolegend, San Diego, CA, USA) and Human IL-8 (CXCL8) Standard ABTS ELISA Development Kit (Peprotech, London, UK), according to the manufacturer’s instructions. The IL-6 and IL-8 amounts were expressed as pg.

NO produced by macrophages was measured using the Griess reaction system (Promega). Mouse IL-6, IL-1β, TNF-α, IFN-γ and GM-CSF level in the cultured medium was determined by Th1/Th2 Cytokine 11-Plex Mouse ProcartaPlex™ Panel according to the manufacturer’s recommendations (Invitrogen). Human IL-6 and TNF-α in culture medium was measured by ELISA MAX Standard Set Human IL-6 (BioLegend, USA) and TNF alpha Human Uncoated ELISA Kit (Invitrogen, USA), respectively, following manufacturer’s instructions.