Human il 1β elisa kit

Following co-culture for 24 h, the concentration of IL-1β in the supernatants was measured using a human IL-1β ELISA kit (MultiSciences Biotech Co., Ltd., Hangzhou, China) according to the manufacturer's protocol. The standards and/or samples were incubated in the wells at 37°C for 1.5 h. Subsequently, the wells were washed and primary antibody was added, covered with an adhesive strip and incubated at 37°C for 1 h. Following washing of the unbound biotinylated antibody, streptavidin-HRP was added for incubation at 37°C for 30 min. A cycle of five washing steps (with PBST for 30 sec each time) was performed and substrate solution was added to the samples. Following incubation at dark for 30 min, the stop solution was added. The OD was detected at 450 nm on a microplate reader (BioTek Instruments, Inc.), and the concentration of IL-1β was calculated according to the manufacturer's protocol.

The concentrations of tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) in the culture medium were detected using a commercial human TNF-α enzyme-linked immunosorbent assay (ELISA) kit (MultiSciences, Hangzhou, China) and human IL-1β ELISA kit (MultiSciences), respectively, according to the manufacturer’s instructions. The activity of NF-κB in cell lysates was detected using the NF-κB p65 Transcription Factor Assay Kit (Abcam, Cambridge, UK) according to the recommended experimental protocol.