Enzyme linked immunosorbent assay (ELISA) for quantification of cytolines (TNFα, IL-6 and IL-10)

In order to measure the cytokine release from SH-SY5Y cells after propofol treatment we used the ELISA kits for quantification of TNFα, IL-6 and IL-10 (product numbers 430208, 430508, 430608 respectively, all BioLegend, San Diego, CA), according to manufacturer’s recommendations. Briefly, cells were treated with 25 μg/mL propofol and left to incubate for 2h, 4h and 24h in complete growth medium. Supernatant of the cells were taken and applied to the ELISA plate. After washing, detection antibodies conjugated to horseradish peroxidase were applied and incubated. After washing away any excess antibodies, we added the HRP substrate and left the reaction for 10 min before stopping it. The resulting colour change was measured at 450 nm.