Mouse il 1β elisa kit

KA and Sodium taurocholate (NAT) was obtained form MedChemExpress (New Jersey, USA). DSPE and PEG₂₀₀₀ were purchased from RuixiBio (Xi’an, China). DIR iodide was offered by Bioss (Beijing, China). Myeloperoxidase (MPO) and Malondialdehyde (MDA) assay kit were acquired from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Mouse IL-1β ELISA Kit and total superoxide dismutase assay Kit were form Beyotime Biotech (Shanghai, China). Antibodies against Bax and Bcl-2 were provided by Proteintech (Wuhan, China). Nrf2 and HO-1were supplied by Bioss (Beijing, China). Caspase3 and Cleaved-Caspase3 were products of Abcam (USA). The SPN-9001 Histostain^™-SP kits was supplied by Zhongshan Jinqiao Biotechnology Co., LTD (Beijing, China). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), if not otherwise stated. All chemicals and reagents were purchased commercially and were used as the manufacturer's instructions.

The concentrations of interleukin- (IL-) 1β, IL-6, and tumor necrosis factor- (TNF-) α in HK-2 cells were detected via the Human IL-1β ELISA Kit (PI305, Beyotime), Human IL-6 ELISA Kit (PI330, Beyotime), and Human TNF-α ELISA Kit (PT518, Beyotime), respectively. The concentrations of IL-1β, IL-6, and TNF-α in kidney tissues were measured via the Mouse IL-1β ELISA Kit (PI301, Beyotime), Mouse IL-6 ELISA Kit (PI326, Beyotime), and Mouse TNF-α ELISA Kit (PT512, Beyotime), respectively.

Cell culture supernatants and lung tissues were collected and frozen at −80 °C. Cytokines were quantified by specific ELISA kits following the manufacturer’s instructions. Human IL-1β (DKW12-3012/-2012), human IL-6 (DKW12-1060/-2060) and human TNF-α (DKW12-1720/-2720) ELISA Kits were purchased from Dakewe Biological Technology Co., Ltd. (Beijing, China). Mouse IL-1β ELISA Kit (PI301), mouse IL-6 ELISA Kit (PI326) and mouse IL-TNFα ELISA Kit (PT512) were purchased from Beyotime Institute of Biotechnology (Shanghai, China). Mouse IL-8 ELISA Kit (MBS261967) were purchased from MyBioSource Bio Co., Ltd. (Vancouver, BC, Canada).

Brain tissue samples were homogenized using RIPA buffer (P0013K, Beyotime, Shanghai, China) and then centrifuged to obtain the supernatant. The levels of IL-1β, IL-6, and TNF-α in the brain were detected by the Mouse IL-1β ELISA Kit (PI301, Beyotime, Shanghai, China), IL-6 ELISA Kit (PI326, Beyotime, Shanghai, China), and TNF-α ELISA Kit (PT512, Beyotime, Shanghai, China), respectively, according to the manufacturer’s protocols. The absorbance was measured at 450 nm using a microplate reader (Multiskan FC, Thermo, Finland).

The levels of IL-1β and TNF-α in the lavage fluid were evaluated using the Mouse IL-1β ELISA kit (Beyotime, PI301) and Mouse TNF-α ELISA kit (Beyotime, PT512). The measurement was carried out in accordance with the product manual.

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.