Human il 1β quantitative elisa kit

Human IL-1β Quantitative ELISA kit (Lot. DLB50, R&D Systems, United States) were used to analyze cell culture supernatant samples according to the manufacturer’s instructions. Briefly, cell culture supernatant samples were equilibrated to the room temperature from stored deep-frozen (−80°C) prior to use. Next, all samples and the diluted standard antibody samples were added into appropriate wells. After incubating for 2 h at room temperature, aspirating the content of every wells and washing well with 400 μL 1 × Washing solution for three times. Then 200 μL human IL-1β Conjugate was added into each well for 1 h at room temperature. After washing for 3 times, adding 200 μL Substrate Solution to all wells for 20 min at room temperature, protecting from light. Last, adding 50 μL of Stop Solution to each well. Determine the optical density of each well within 30 min at 450 nm.