Enzyme-linked Immunosorbent assay for IL-6, TNF-α, IFN-γ, IL-17 and IL-22
The cytokine levels in the serum were collected and allowed to clot at 4 °C for 2 hours. The clots were removed by centrifuging at 7000 rpm for 30 min in a refrigerated centrifuge. Serum was obtained from all subjects by centrifugation and stored at-80°C for determination of cytokines levels. The levels of cytokines in the serum/supernatant were measured by Enzyme-linked Immunosorbent Assay (ELISA) using commercially available IL-6, TNF-α, IFN-γ, IL-17 and IL-22 ELISA kits (BOSTER, Wuhan, China) according to the manufacturer’s instructions.