Opteia mouse elisas

To analyze the levels of IFN-γ, IL-12p40, IL-4, IL-6, and TNF-α in lung tissue, OptEIA mouse ELISAs were purchased from BD Biosciences. IL-13 levels were assessed using the AbFrontier Cymax mouse ELISA kit (AbFrontier, Seoul, Korea). All assays were conducted according to the manufacturers’ guidelines. All lung samples were prepared by lysis buffer made with a protease inhibitor cocktail and RIPA buffer (Thermo Fisher Scientific). Aliquots of lung tissue from all groups were weighed and homogenized with lysis buffer. They were centrifuged at 8200 rpm for 15 min, and the supernatants were harvested and measured using a microplate reader (EZ Read 400, Biochrom, Cambourne, UK).

ELISA was conducted following the methods of our previous study (Lee et al., 2019 (link)). Whole proteins were collected after cell lysis from the lung tissue using a buffer to measure the levels of proteins such as IFN-γ, IL-5, IL-13, and TNF-α. The buffer was made with a protease inhibitor and RIPA buffer (Thermo Fisher Scientific, Waltham, MA, USA). OptEIA mouse ELISAs (BD Biosciences, San Jose, CA, USA) were used to evaluate the protein levels of IFN-γ, IL-5, and TNF-α. IL-13 measurements were taken using the AbFrontier Cymax mouse ELISA kit (AbFronteir, Seoul, Korea) according to the manufacturer’s instructions. Whole proteins of identical weights from each group were homogenized with a lysis buffer and the supernatants were collected and analyzed with a microplate reader after centrifugation (MUTISKAN Sky, Thermo Fisher Scientific).