Sta liquid fib

The plasma concentration of fibrinogen and D-Dimer was quantitatively determined using the STA-Liquid fib and the Liatest D-Di kits (Diagnostica Stago, Asnieres-sur-Seine, Paris, France), respectively, and a STA-R coagulation analyser (Diagnostica Stago). In-house prepared ELISAs employing specific mAbs were used to determine the plasma concentration of tPA and PAI-1 [31 (link)].
The rate of fibrin formation (Vmax) and fibrin structure measurements (mass-length ratio [(ML), diameter (Diam) and density (Dens)] and fibrin clot lysis were determined according to Sjøland et al.[32 (link)] using turbidity measurements
Fibrin clot lysis was measured by mixing 60 μl of plasma with 120 μl of a reaction mixture consisting of 1 IU/ml of thrombin, 15 mmol/l CaCl₂, 50 mmol/l Tris-HCL buffer, 150 mmol/l NaCl and 0.6 mg/ml recombinant tPA (Boehringer Ingelheim, Ingelheim am Rhein, Germany). The optical density at 340 nm (OD340) was recorded for 30 min, and the percentage of the clot lysed thereafter was determined. Measures of fibrin clot properties was determined using a similar setup, but replacing rtPA with Tris-HCl buffer. The fibrin fibre properties (i.e. Vmax, mass-length, Diam and Dens) were determined by measurements of OD405, OD560, OD608 and OD690 of the fibrin clot and calculated according to Carr et al.[33 (link),34 (link)].