On the reliability of the use of heparin immobilized on agarose for the study of the interactions among heparin, thrombin and antithrombin.
The extent of inhibition of thrombin was re-examined as a consequence of the sequence of addition of thrombin and antithrombin III to a column of heparin immobilized on agarose. With the use of pure enzyme, pure inhibitor, and a highly sensitive chromogenic substrate, no appreciable difference in the extent of thrombin inhibition was observed between the two sequences of addition. These observations, together with a demonstrated sensitivity of the method to variations in experimental protocol, challenge the conclusions reached in an earlier work (Hatton and Regoeczi, Thromb. Res. 1977; 10:645) which utilized this experimental approach but which employed larger quantities of reactants and a less sensitive substrate.
D000818:Animals; D000991:Antithrombins / Q000378:metabolism*; D001780:Blood Coagulation Tests / Q000379:methods; D002417:Cattle; D002847:Chromatography, Agarose; D006493:Heparin / Q000378:metabolism*; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000991:Antithrombins; D006493:Heparin; D013917:Thrombin