Legend maxtm elisa kits with pre coated plates

Legend Max^TM ELISA kits with pre-coated plates (BioLegend UK Ltd) were used to measure the pro-inflammatory cytokine IL-1β and anti-inflammatory cytokines, IL-10 and TGF-β, present in the cell culture supernatant of the different conditions tested following the manufacturer’s procedure.

To characterize the cytokine level profile in response to immunization and prior to the challenge, we determined the concentrations of IFN-γ, IL-2, IL-4, IL-10, and IL-12p70 in splenocyte culture supernatants using Legend Max^TM ELISA Kits with Pre-coated Plates (Biolegend, San Diego, California, USA). Splenocytes obtained from immunized mice and their control counterparts at two weeks post final immunization were cultivated in 96-well plates. The splenocyte culture supernatants were collected at 24 h for the measurement of IL-2 and IL-4, at 72 h for the measurement of IL-10, and at 96 h for the measurement of IFN-γ and IL-12p70. All samples were tested in triplicate.