Mouse il 1β elisa max standard

The protein concentrations of the intestinal samples were measured by Bradford’s method using BSA as the protein standard. Inflammatory cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor-alpha (TNF-α) in the intestinal samples were tested using enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer instructions (mouse IL-1β ELISA MAX Standard [Biolegend, San Diego, CA, USA], mouse IL-6 and TNF-α ELISA, [Thermo Fisher Scientific, Rockford, IL, USA]). Briefly, samples were added to the overnight coated 96-well plates with IL-1β, IL-6, and TNF-α capture antibodies and incubated for 1 h. at room temperature (RT), washed four times, and then incubated with diluted detection antibody solution. After incubation with detection antibodies, the plate was washed four times with a washing buffer, and then incubated with horseradish peroxidase-linked streptavidin solution for 30 min at RT in the dark, and finally incubated with diluted substrate solution. The absorbance was measured at 420 nm by a microplate reader (Byoany Absorbance 96, Germany).