FXIIIa cross-linking of BβD432A and normal recombinant fibrinogen. Cross-linking was examined by 8% SDS-PAGE under reducing conditions. Fibrinogen (5 μg) was incubated with either FXIIIa and thrombin at final concentrations of 3.3 U/mL and 0.07 U/mL, respectively (A: normal, B: BβD432A) or FXIIIa and batroxobin at final concentrations of 3.3 U/mL and 0.08 U/mL, respectively (D: normal, E: BβD432A). The samples were incubated for 0 minutes (lane 1), 2 minutes (lane 2), 5 minutes (lane 3), 10 minutes (lane 4), 20 minutes (lane 5), 40 minutes (lane 6), 1 hour (lane 7), and 2 hours (lane 8). Data were analyzed by determining the ratio of γ-γ dimers to Bβ-chain by densitometry and are plotted against time (thrombin-polymerized; C: ■ indicates normal; □, BβD432A; and batroxobin-polymerized: F: ● indicates normal; ○, BβD432A). The error bars (C,F) represent SDs of 3 experiments.