Legend maxtm kits

TNF-α, IL-4, IL-17A, IFNγ, IL10, IL22, TGFβ, and IL23 were accurately quantified in duplicate using sandwich enzyme-linked immunosorbent assay (ELISA) kits purchased from BioLegend Inc., USA (LEGEND MAXTM kits). The normal concentrations of TNF-α, IL-4, IL-17A, IFNγ, IL10, IL22, TGFβ, and IL23 were 3.5–200 pg/ml, 0.6–250 pg/ml, 0.8–1,000 pg/ml, 5.6–1,000 pg/mL, 3.15–200 pg/mL, 31.3–2,000 pg/mL, 0.23–100 pg/mL, 15.6–2,000 pg/mL, respectively. The variability of the duplicates was <10% of the mean value. According to the manufacturer's instructions and each assay with a standard curve, the values were recorded at wavelengths of 450 and 570 nm and were then calculated from the standard curve. The TNF-α, IL-4, and IL-17A ELISA results had intra-assay CVs of 2.3–10.8%, 4.5–5.3%, 5.4–5.6% and inter-assay CVs of 5.0–6.7%, 6.9–7.1%, 7.8–9.1%, respectively.