Ab222503

Enzyme linked immunosorbent assay (ELISA) was used to further confirm the expression of above inflammation related cytokines before and after SAH. The process of measurement was as per the manufacturer’s protocol and previous studies (Li et al. 2020 (link); Zhang et al. 2019a (link), b (link)). The left hemisphere cortex tissue suspension and protein samples were acquired similar to the WB method. The protein samples and the standard were mixed with antibody cocktail in pre-coated 96-well microplate, and then incubated at room temperature (25 ℃) for 1 h. After washing with buffer reagent, the development solution was added to each well, and finally the stop solution was added. Using a spectrophotometer to analyze the optical density (OD) at 450 nm for each well in the microplate, the protein concentration could be quantified by comparing the OD of target protein with standard curve. ELISA kits in the study were used: IL-1β (ab197742, Abcam, USA), IL-6 (ab222503, Abcam, USA) and TNF-α (ab208348, Abcam, USA).

Mice were anesthetized and were transcranial perfused with 50 mL of cold NS. The temporal cortex was collected for assessment. The concentrations of proinflammatory cytokines were detected. There were five supernatants in each group for ELISA analysis. An ELISA was used as directed by the manufacturer to determine the levels of IL‑1β (cat. no. ab197742; Abcam), IL‑6 (cat. no. ab222503; Abcam), TNF‑α (cat. no. ab208348; Abcam), and NF‑κB (cat. no. ab176663; Abcam). The specific experimental procedure according to the guidelines stipulated by the manufacturer and previous study [45 (link)].