F1.2 and TAT were measured using Enzygnost F1.2 (monoclonal) and Enzygnost TAT micro enzyme-linked immunosorbent assay kits (Siemens Healthcare Diagnostics, Marburg, Germany). The fibrin fragment D-dimer was measured using the Innovance D-dimer kit and a BCS XP coagulation analyzer (both Siemens Healthcare Diagnostics). All analyses were performed according to the manufacturer's instructions. Levels of thrombin and APC were measured using the thrombin– and APC–OECA as previously described.
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Thrombin generation kinetics were measured using calibrated automated thrombography (CAT) (Thrombinoscope BV, Maastricht, the Netherlands) using standard reagents (Stago, Düsseldorf, Germany) and a Fluoroskan Ascent FL plate reader (Thermo Scientific) as described elsewhere.
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Reference ranges for all parameters were previously established by analyzing plasma samples of healthy individuals. Normal plasma levels of thrombin and APC were found below the lower limit of quantification (LLOQ) of the assays.
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Thus, the LLOQ was defined as the upper limit of the reference range of these parameters.