FIG. 4.

Binding efficiency of CHB3 in dependence on the pH. As described in Materials and Methods, CHB3 was incubated with ground crab shell powder (α-chitin) at room temperature overnight in buffers adjusted to pH 5 (lanes 1 and 5), pH 6 (lanes 2 and 6), pH 7 (lanes 3 and 7), and pH 8 (lanes 4 and 8). After centrifugation, the supernatants were analyzed by SDS-PAGE (lanes 1 to 4). The pellets were washed with the corresponding buffer, resuspended, boiled in the SDS-PAGE sample buffer for 10 min, and centrifuged. The supernatant was subjected to gel electrophoresis (lanes 5 to 8). CHB3 was detected by immunoblotting with anti-CHB1 antibodies, and the relative quantities were determined by scanning. The highest amount of cross-reacting protein was set as 100%. The sizes of the protein markers are indicated.