Mouse il 18 elisa kit

Sample preparation: Tears were collected from the lateral canthus of mice using glass microcapillary tubes. One sample consisted of tears from both eyes of one mouse that were pooled in phosphate buffered saline (PBS) + 0.1% BSA and stored at −80°C until the assay was performed (Dogru et al., 2022 (link)). Hippocampus tissues of mice were isolated, snap-frozen in liquid nitrogen, and stored at − 80°C for protein extraction. The tissues rinsed with cold PBS to remove residual blood, weighed, and added with PBS containing protease inhibitor cocktail (1:9 w/v). Samples were then homogenized, centrifuged at 3500 ×g for 20 min, and supernatants were collected (Xue et al., 2021 (link); Dang et al., 2022 (link)).
The levels of inflammatory cytokines including tumor necrosis factor (TNF)-α, IL-1β, and IL-18 in hippocampi and tears were detected using Mouse TNF-α ELISA Kit (PT512, Beyotime), Mouse IL-1β ELISA Kit (PI301, Beyotime), and Mouse IL-18 ELISA Kit (PI553, Beyotime), respectively, as per company instructions.

The concentration of IL‐1β in cell culture supernatant was detected with Mouse IL‐1β ELISA Kit (catalogue number: PI301, Beyotime), while the concentration of TNF‐α, IL‐6, IL‐18 and MCP‐1 in both BALF and cell culture supernatant were examined by Mouse TNF‐α ELISA Kit (catalogue number: PT512, Beyotime), Mouse IL‐6 ELISA Kit (catalogue number: PI326, Beyotime), Mouse IL‐18 ELISA Kit (catalogue number: PI553, Beyotime) and Mouse CCL2/JE/MCP‐1 ELISA Kit (catalogue number: PC125, Beyotime) in keeping with the working instructions. The absorbance was determined with a microplate reader (Thermo Fisher Scientific) at 450 nm.