Il 6 enzyme linked immunosorbent assay kits

BMCMCs (5 × 10⁶) were incubated with or without yeasts (9.4 × 10⁷, 4.7 × 10⁷, or 2.35 × 10⁷) of S. schenkii or S. albicans at 37°C in supplemented RPMI 1640 medium under an atmosphere of 5% CO₂ for 24 h. Cell-free supernatants were collected and stored at −80°C until assayed for TNF and IL-6. Then, we used ELISA kits (Biolegend) to determine the levels of TNF, IL-6, IL-1β, IL-10, and MCP-1 in cell culture media according to the manufacturer's instructions. Serum levels of TNF and IL-6 in sporotrichosis patients were determined using human TNF or IL-6 enzyme-linked immunosorbent assay kits, respectively (R&D Systems). The results were expressed in pg/ml.

After treatment, the levels of IL-1β, TNF-α, and IL-6 in the renal tissues were measured by using IL-1β, TNF-α, and IL-6 enzyme-linked immunosorbent assay kits (R&D Systems, Minnesota, USA), as per manufacturer instructions.