Detection of bovine inflammatory cytokines IL-1?, IL-6, and TNF-? with a multiplex electrochemiluminescent assay platform

Commercially available bovine-specific assays are limited in number, and multiplex for this species rare. Our objective was to develop a assay the bovine inflammatory cytokines IL-1?, IL-6, TNF-? using Meso Scale Discovery U-PLEX platform. "Do-It-Yourself" ELISA kits that contained polyclonal antibodies, both unlabeled biotinylated, specific recombinant cytokine standard, were purchased each of these three cytokines. The biotinylated antibodies coupled linkers bind locations within well plate. Unique used conjugated with electrochemiluminescent labels serve as detection antibodies. Each optimized individually prior performing an optimization on containing reagents all To calculate concentrations, standard curves developed run concurrently Standard IL-1? at concentrations ranging from 0 50,000 pg/mL, IL-6 10,000 pg/mL. average lowest level concentration measured by 5.3 0.92 22.34 pg/mL respectively, determined data seven plates plasma samples combination healthy diseased cattle. platform viable means custom analyte- species-specific privately or sets commercially reagents.