FIGURE 1.

Characterization of PNGase F-treated APC. A, APC and PNGase F-treated APC (PNG-APC, 1 μg) were characterized by 4–15% SDS-PAGE analysis and Western blotting, using a sheep anti-protein C/APC polyclonal antibody that detects the protein C/APC heavy chain (HC, shown). B, the anticoagulant activity of APC (○) and PNG-APC (■ 2.5–20 nm) was assessed in protein C-deficient plasma by a thrombin generation assay. Thrombin generation was initiated with 5 pm soluble tissue factor and 100 mm CaCl₂ and assessed as described under ”Experimental Procedures.“ The percentage of ETP was determined for thrombin generation in the presence and absence of APC, with 100% ETP defined as ETP in the absence of either APC species.