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Solid-phase-heparin as heparin-agarose [Hep-Ag] was used for the isolation of the complex formed by antithrombin and thrombin. This complex has a lower affinity to heparin than antithrombin III. The complex has no antithrombin activity. The increase of the complex during blood coagulation can be seen by the two-dimensional Immunoelectrophoresis in agaroee, which contains heparin (Sas et al., Thromb.Res. 6, 87-91, 1975).  Platelet factor 4 (PF 4) was isolated by a three step procedure: 1. precipitation by divalent cations according to Burstein, 2. adsorption to Hep-Ag and desorption with high salt concentrations, 3. desalting over Sephadex G 25 fine. PF 4 was prepared from isolated platelets, platelet rich plasma and serum.

Isolation of the Antithrombin-Thrombin Complex and Platelet Factor 4 by Solid Phase Heparin