Human il 1 beta il 1f2 quantikine elisa kit

The amount of IL-1β in gingival crevicular fluids and cell culture supernatants from different groups was measured by ELISA according to the manufacturer’s guidelines. A Human IL-1 beta/IL-1F2 Quantikine ELISA Kit (R&D, United States) was used.

Rat primary cultured microglia (2 × 10⁵ cells/well) were cultured in 24-well plates for 1 h, after which the cells were incubated for 1 h with fresh serum-free medium. LPS (500 ng/mL for 3 h)-primed microglial cells were treated with compounds (1–10 μM for 30 min) and then stimulated with ATP (2 mM for 30 min, co-treated with each compound). PMA-differentiated THP-1 cells (2 × 10⁵ cells/well) were cultured in 24-well plates for 24 h. LPS (500 ng/mL for 3 h)-primed cells were treated with compounds (10 μM for 30 min) and then stimulated with BzATP (300 μM for 30 min, co-treatment with each compound). The supernatants were used to measure the IL-1β release. The concentration of IL-1β in each sample was determined using the Rat IL-1 beta/IL-1F2 Quantikine ELISA Kit (RLB00, R&D Systems, Minneapolis, MN, USA) or Human IL-1 beta/IL-1F2 Quantikine ELISA Kit (DLB50, R&D Systems, Minneapolis, MN, USA). Each assay was performed in accordance with the manufacturer’s instructions.

Culture supernatant was collected from 24-well plates and concentrations of MCP-1, IL-1β, IL-6, and TNF-α were measured by human CCL2/MCP-1 Quantikine ELISA kit, human IL-1 beta/IL-1F2 Quantikine ELISA kit, human IL-6 Quantikine ELISA kit, and human TNF-alpha Quantikine ELISA kit (all from R&D Systems, UK), respectively, according to the manufacturer's instructions.