Elisa max deluxe set human il 6 kit

THP-1 cells were seeded at \(3 \times {10}^{5}\) cells/well in 24-well microplates and differentiated into macrophages by treatment with 50 nM of phorbol 12-myristate 13-acetate (Sigma-Aldrich), for 48 h. Cells were exposed to the fermentations extracts (10 and 1 mg.mL -1 ) for 24h, in the presence or absence of lipopolysaccharides form E. coli O111:B4 (LPS, Sigma-Aldrich) to induce inflammation. For anti-inflammatory control, macrophages were treated with 20 nM of betamethasone (Sigma-Aldrich). After 24h, supernatants were collected and the level of proinflammatory cytokine IL-6 was determined by ELISA, using the ELISA MAX™ Deluxe Set Human IL-6 kit (Biolegend), according to manufacturer's instructions. For total protein quantification, cells were lysed with water, and BCA method was performed, as previously described. The results were expressed in pg of cytokine.ug -1 of total protein.