Figure 3. Cross-linking of semenogelin. (A) Gel stained with Coomassie Brilliant Blue. (B) Gel photographed in ultraviolet light. Lanes 1–3 respectively represent rSgI, SgI₄₃, and SgI (not crosslinked). Lanes 4–9 show duplicates of rSgI, SgI₄₃, and SgI, each at a final concentration of 2 µmol/L. To achieve cross-linking, 5 nmol thrombin, 40 mmol/L DTT, 40 µg/mL FXIIIa, 0.7 mmol/L MDTC (mono-dansyl-thio-cadaverine), and finally 10 mmol/L CaCl₂ in 50 mmol/L TBS (pH 7.5) were added to each of the semenogelin samples, which were subsequently incubated for 2 hours at 37°C. The reaction was terminated with 20 mmol/L EDTA, and 15 µL of the reaction solution was applied on the gel.