FIG. 5.

Effect of CAP18_109–135 peptide on the Limulus assay. In experiment 1, LPS (50 pg/ml) was mixed with CAP18_109–135 (400 μg/ml) and incubated at 37°C for 10 min. Then 50 μl of the Endospecy reagent was added to the mixture, which was incubated at 37°C for a further 30 min. At the end of the incubation, the absorbance of the reaction mixture was measured at 405 nm (A₄₀₅). In experiment 2, (1 → 3)-β-d-glucan (48.4 pg/ml in 25 μl) was incubated with CAP18_111–137 (200 μg/ml in 25 μl) at 37°C for 60 min. Then G-test reagent was added to the mixture, and the mixture was incubated at 37°C for a further 30 min. At the end of the incubation, the absorbance of the reaction mixture was measured at 405 nm. In experiment 3, LPS (10 μg/ml) was mixed with a solution of coagulation factors prepared from horseshoe crab (factor C, factor B, and proclotting enzyme) and the mixture was incubated at 37°C for 30 min. Then CAP18_111–137 (315 μg/ml) was added to the mixture, which was incubated at 37°C for a further 60 min. Finally, the substrate solution was added to the mixture, and the mixture was incubated at 37°C for a further 30 min. At the end of incubation, the absorbance of the reaction mixture was measured at 405 nm.