American Society of Hematology

Figure 3

$Figure 3. Trypsin digestion of VWF. (A) Twelve percent SDS-PAGE analysis of the major fraction obtained by ion-exchange chromatography of trypsin-digested VWF under reducing (R) and nonreducing (NR) conditions followed by Coomassie staining. (Vertical line indicates a repositioned gel lane.) (B,C) Of the purified 55-kDa fragment, 20 nM was partially digested with PNGase F for 2 hours at 37°C (B) and completely digested with PNGase F overnight at 37°C (C), and analyzed in 12% SDS-PAGE gels followed by Western blotting to nitrocellulose membranes and probing with anti–VWF-HRP. (D) Partially PNGase F–digested VWF 55-kDa fragment was also detected with elderberry bark lectin (EBL) specific for terminal sialic acid residues (left panel) and Ulex europaeus specific for the H-antigen (right panel).$

Trypsin digestion of VWF. (A) Twelve percent SDS-PAGE analysis of the major fraction obtained by ion-exchange chromatography of trypsin-digested VWF under reducing (R) and nonreducing (NR) conditions followed by Coomassie staining. (Vertical line indicates a repositioned gel lane.) (B,C) Of the purified 55-kDa fragment, 20 nM was partially digested with PNGase F for 2 hours at 37°C (B) and completely digested with PNGase F overnight at 37°C (C), and analyzed in 12% SDS-PAGE gels followed by Western blotting to nitrocellulose membranes and probing with anti–VWF-HRP. (D) Partially PNGase F–digested VWF 55-kDa fragment was also detected with elderberry bark lectin (EBL) specific for terminal sialic acid residues (left panel) and Ulex europaeus specific for the H-antigen (right panel).

This Feature Is Available To Subscribers Only