Fig. 7.
Formation of A2-FVIII337-372 cross-linked product. A2 (200 nmol/L) was incubated with no additions (lane 1), 0.625 μmol/L (lane 2), 1.25 μmol/L (lane 3), 2.5 μmol/L (lane 4), 5 μmol/L (lane 5), 10 μmol/L (lane 6), or 20 μmol/L (lane 7) FVIII337-372 in a reaction volume of 20 μL for 30 minutes at room temperature. EDC (200 μmol/L) was added and incubation was continued for 1 hour. After addition of SDS sample buffer, reactions were subjected to electrophoresis on 12% gels, transferred to PVDF, and immunoblotted with R8B12 (A) or anti-FVIII337-372 (B).

Formation of A2-FVIII337-372 cross-linked product. A2 (200 nmol/L) was incubated with no additions (lane 1), 0.625 μmol/L (lane 2), 1.25 μmol/L (lane 3), 2.5 μmol/L (lane 4), 5 μmol/L (lane 5), 10 μmol/L (lane 6), or 20 μmol/L (lane 7) FVIII337-372 in a reaction volume of 20 μL for 30 minutes at room temperature. EDC (200 μmol/L) was added and incubation was continued for 1 hour. After addition of SDS sample buffer, reactions were subjected to electrophoresis on 12% gels, transferred to PVDF, and immunoblotted with R8B12 (A) or anti-FVIII337-372 (B).

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