The following reagents were from Sigma–Aldrich (St. Louis, MO, USA); 16-mercaptohexadecanoic acid, cysteamine, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, succinic anhydride and N-hydroxysuccinimide. Triethylamine, dimethylformamide and bovine serum albumin (BSA) were from Fisher Scientific (Pittsburgh, PA, USA). Six-armed-poly(ethyleneglycol)–amine was from SunBio (South Korea). PBS (Cat#70011-036) and Tween-20 were from Life Technologies (Grand Island, NY, USA).
Sandwich antibody pairs for IL-20 were produced by Novo Nordisk A/S. Capture anti-IL-20 antibody was of the IgG1 isotype. Unspecific binding was assessed using a IgG1 isotype control from R&D Systems (MAB002, Minneapolis, MN, USA). IRDye-800 streptavidin conjugate was from LiCor Biosciences (Lincoln, NE, USA). Mouse, goat and bovine IgG were from Jackson ImmunoResearch (West Grove, PA, USA).
Purified cytokine antigen standards and sandwich antibody pairs for IL-1β (MAB601, BAF201), IL-10 (MAB2172, BAF217), IL-6 (MAB206, BAF206), and TNFα (MAB610, BAF210) were purchased from R&D systems. All capture antibodies were of the IgG1 isotype.
Sandwich antibody pairs for IL-20 were produced by Novo Nordisk A/S. Capture anti-IL-20 antibody was of the IgG1 isotype. Unspecific binding was assessed using a IgG1 isotype control from R&D Systems (MAB002, Minneapolis, MN, USA). IRDye-800 streptavidin conjugate was from LiCor Biosciences (Lincoln, NE, USA). Mouse, goat and bovine IgG were from Jackson ImmunoResearch (West Grove, PA, USA).
Purified cytokine antigen standards and sandwich antibody pairs for IL-1β (MAB601, BAF201), IL-10 (MAB2172, BAF217), IL-6 (MAB206, BAF206), and TNFα (MAB610, BAF210) were purchased from R&D systems. All capture antibodies were of the IgG1 isotype.