Figure 3

Further mapping of the C9 binding site in overlapped synthesized peptides. Different overlapped peptides within the C9 binding fragment of Ts-Pmy (Pmy831-885) were synthesized, and then spotted onto a nitrocellulose membrane by using a narrow-mouth pipette tip (5 μg in 2.5 μl). After being blocked with 1% BSA in PBS, the membrane was incubated with human C9 (1 μg/ml) at 37°C for 2 h and then with anti-C9 mAb (0.2 μg/ml) at room temperature for 1 h. IRDye 800CW-labeled goat anti-mouse IgG (50 ng/ml) was used as the secondary antibody. The peptide size diagram is shown on the left and the Dot-blot with C9 is shown on the right.