Electrophoretic analysis of VWF constructs dA1, dA3, and dA13.

Recombinant normal human VWF (WT) and variants with deletions of dA1, dA3, or both dA1 and dA3 (dA13) were expressed in transiently transfected COS-7 cells, and conditioned medium was collected for analysis. (A) The multimer distribution of plasma vWF (plasma) and recombinant VWF preparations were compared by SDS–1.5% agarose gel electrophoresis under nonreducing conditions. The position of the smallest multimer, an approximate 500 kd dimer, is indicated. (B) VWF subunits and a 200 kd standard protein (M) were analyzed by SDS-PAGE on a 5% polyacrylamide gel under reducing conditions. The masses of the subunits are about 250 kd (WT), about 225 kd (dA1 or dA3), and about 205 kd (dA13).