Figure 7.

Evidence of binding of S100A8/A9 with proinflammatory cytokines. For detection of S100A8/A9–proinflammatory cytokine complexes, we used the ELISA plate coated with anti-S100A8/A9 monoclonal IgG (Mo2B9). Biotinylated anti-rat IL-1β, IL-6, and TNF-α IgGs were used as the second antibody. Finally, colour development was achieved by measuring horseradish peroxidase activity after incubation of streptavidin–horseradish peroxidase conjugate for 30 min. The reaction was significantly positive and quantitative, indicating the existence of S100A8/A9–cytokine complexes. This is a representative result of three samples.