Tans G, Janssen-Claessen T, Rosing J
Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
Thromb Haemost. 1989 Jun 30;61(3):386-91.
In this paper we report a method via which enzymatically active products formed during prothrombin activation can be detected by simple photographic means after SDS-gel electrophoresis, blotting onto a nitrocellulose membrane and visualization with the chromogenic substrate, S2238. After amidolytic detection the same nitrocellulose membrane can also be used for immunologic detection of prothrombin activation products, thus allowing a complete description of product formation during prothrombin activation. The detection limit of the so-called "amidoblot" is approximately 3 ng thrombin per gel sample which is comparable to the sensitivity of immunoblotting. It is further shown that the amidoblot technique can also be applied to other coagulation factors for which a suitable chromogenic substrate is available (factor XIIa, kallikrein, factor XIa, factor Xa, plasmin and activated protein C).
在本文中，我们报告了一种方法，通过该方法，凝血酶原激活过程中形成的具有酶活性的产物在十二烷基硫酸钠 - 凝胶电泳、转印至硝酸纤维素膜并用显色底物S2238显色后，可通过简单的照相手段进行检测。在酰胺分解检测之后，同一硝酸纤维素膜还可用于凝血酶原激活产物的免疫检测，从而能够完整描述凝血酶原激活过程中的产物形成情况。所谓的“酰胺印迹法”的检测限约为每个凝胶样品3 ng凝血酶，这与免疫印迹法的灵敏度相当。进一步表明，酰胺印迹技术还可应用于有合适显色底物的其他凝血因子（因子XIIa、激肽释放酶、因子XIa、因子Xa、纤溶酶和活化蛋白C）。
