Verikine human ifn α elisa kit

Archived samples (never thawed, stored at −80 °C) were used for all cytokine measurements. Levels of IFN-α were determined in the serum using a high-sensitivity VeriKine human IFN-α ELISA Kit (PBL Assay Science, Piscataway, NJ, USA), which detects all 12 human IFN-α subtypes (dynamic range 1.95–125 pg/mL), and were also quantified using a Simoa HD-1 instrument with the Simoa® IFN-α Advantage Kit HD-1/HD-X (Quanterix Corp., Billerica, MA, USA), which has a linear range from 0 to 60 pg/mL and an LLOQ = 0.016 pg/mL. Levels of Galectin-9 and CXCL-10 (IP-10) were measured in diluted serum (1/5 dilution) using DuoSet ELISA kits (R&D Systems, Minneapolis, MN, USA), with dynamic ranges of 93.8–6000 pg/mL and 31.2–2000 pg/mL, respectively. The levels of IFN-γ in neat serum were determined using a Quantikine High Sensitivity ELISA kit (R&D Systems), following the manufacturer’s instructions (dynamic range 0.5–30 pg/mL). Except for some of the Simoa assays, all measurements were performed in duplicate.

IL-1β and IFN-α secretion in supernatants from cell cultures were analyzed using a Duo Set ELISA kit (R&D Systems) and a VeriKine human IFN-α ELISA kit (PBL assay science, NJ, US), respectively, both according to manufacturer’s instructions.

Standard ELISA methods were used to measure concentrations of Gal-9 (R&D Systems), cytokines and chemokines such as IL-12p40, IL-12p70, MIP-1β (macrophage inflammatory protein [MIP]), monocyte chemoattractant protein 2 (MCP-2), tumour necrosis factor-alpha (TNF-α), IL-6, transforming growth factor-beta1 (TGF-β1), IL-18, IL-23, IL-27, and IL-1β (R&D Systems) and IFN-α (Verikine™ Human IFNα ELISA Kit, PBL Assay Science, Piscataway, NJ, USA).