Human il 18 il 1 f4 elisa

IL-1α, IL-1β and IL-18 productions in HaCaT cells were evaluated by using human IL-1alpha Platinum ELISA (BMS243/2, eBiosciences), human IL-1 β/IL-1F2 Quantikine HS ELISA (HSLB00C, R&D systems), human IL-18/IL-1F4 ELISA (7620, R&D systems) kit, respectively. For dose-dependent assay, cells were plated in 12-well plates at a density of 1×10⁵ cells/well and treated with 0.5, 2 or 10 µM sodium arsenite in DMEM medium for 24 h and the media were collected and analyzed. For time dependent assay, cells were exposed to 2 µM sodium arsenite for 24, 48 or 72 h, respectively. The media were collected for ELISA assay. IL-1α, IL-1β and IL-18 productions in the mouse skin were evaluated by using mouse IL-1alpha Platinum ELISA (BMS611, eBioscience), mouse IL-1 β ELISA Kit (ELM-ILb-CL, Rabiotech), mouse IL-18 platinum ELISA kit (BMS618/3, eBioscience), respectively. Skin tissues were harvested and homogenized in extraction buffer (containing 10 mM Tris PH7.4, 150 mM NaCl, 1% Triton X-100). The homogenates were centrifuged at 10,000×g for 20 min at 4°C and the supernatants were used for the determination of cytokine levels by ELISA assay per the manufacturer's instruction. All the assays were done in triplicate. The adjusted quantities were presented as an average of the data ± standard error of mean (SEM).