Max deluxe sets

TNF-α, IL-1β, IL-6, IL-22 and granulocyte/macrophage colony-stimulating factor (GM-CSF) concentrations were determined in serum aliquots, while those of IL-10 and IL-22 were determined in serum aliquots and hepatic lysates. Hepatic IL-17A concentration was also quantified. The concentrations of these cytokines were determined by ELISA kits (MAX™ Deluxe Sets, BioLegend, United States). Briefly, portions from the liver (10% w/v) were homogenized and lysed in an ice cooled lysis buffer (10 mM Tris pH 7.4, 150 mM NaCl, 0.5% Triton X-100) containing a protease inhibitor cocktail (Roche Diagnostics, Germany). Thereafter, the lysed liver aliquots were centrifuged at (5,000 g, 10 min, 4 °C), and the clear supernatants were loaded on a 96-well plate precoated overnight with the appropriate capture antibody. Serum aliquots were prediluted in physiological saline (2:1), followed by loading on the 96-well plate like those of lysates. The routine protocol steps of ELISA were then followed as provided by the manufacturer. The concentration of protein was also assessed in the liver lysates by the Bradford’s assay (Bradford, 1976 (link)).