Human il 10 elisa kit

The inflammatory cytokines including tumor necrosis factor alpha (TNF‐α), interleukin‐1 beta (IL‐1β), interleukin‐10 (IL‐10), and interleukin‐17 (IL‐17) in the plasma were determined by Enzyme‐linked immunosorbent assay (ELISA) with ELISA kits, including Human TNF‐α ELISA Kit (Thomas, KHC3014), Human IL‐1β ELISA Kit (Thomas, BMS224HS), Human IL‐10 ELISA Kit (Abcam, ab100549), and Human IL‐17 ELISA Kit (Abcam, ab83707), which were ready‐to‐use, according to the instructions of manufacturer. In brief, firstly, samples or standards were added to the 96‐well plates, followed by the antibody mix. After incubation, the wells were washed to remove unbound material. TMB substrate (tetramethylbenzidine, TMB) was added, generating blue coloration. This reaction was then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal was generated proportionally to the amount of bound analyte (Biotek), and the intensity was measured at 450 nm. Each reaction was run in triplicate by the same operator. Standard curves were prepared before the antibody reaction, and the standard curve range of Human TNF‐α ELISA Kit (Thermo, KHC3014), Human IL‐1β ELISA Kit (Thermo, BMS224HS), Human IL‐10 ELISA Kit (Abcam, ab100549), and Human IL‐17 ELISA Kit (Abcam, ab83707) were 0.5‐32pg/mL, 0.16 ‐10 pg/mL, 2.34‐150 pg/mL, and 3.12‐100 pg/mL, respectively.