Mouse tnf α elisa maxtm standard set

The concentrations of pro-inflammatory cytokines, including IL-6 and TNF-α, in the resected mouse colon tissues from the experimental groups were evaluated using Mouse IL-6 ELISA MAXTM Deluxe Set and Mouse TNF-α ELISA MAXTM Standard Set (Biolegend, San Diego, CA, USA), respectively. For sample preparation, frozen colon tissue specimens were homogenized in a lysis buffer solution (10 mL of 1 M tris-hydrochloric acid adjusted to pH 8.0) containing a protease inhibitor cocktail. When the colon specimens were homogenized, the solution was centrifuged for 5 min at 3300× g. The supernatant and pro-inflammatory cytokine concentrations were then assayed using an ELISA kit, according to the manufacturer’s instructions.

Supernatants of stimulated cells were harvested and analyzed for IL-6 and TNF-α using mouse IL-6 ELISA MAXTM Standard Set and mouse Tnf-α ELISA MAXTM Standard Set, respectively (BioLegend, San Diego, CA, USA). A TecanGENios Pro plate reader (Tecan, Crailsheim, Deutschland) was used for quantification. Results were analyzed using the Magellan5 software.