An Antithrombin Reaction Related to Prothrombin Activation
An Antithrombin Reaction Related to Prothrombin Activation
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DOI：
10.1152/ajplegacy.1953.176.1.97
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发表时间：
1953-12
影响因子：
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通讯作者：
W. Seegers;J. F. Johnson;C. Fell
中科院分区：
文献类型：
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作者：
W. Seegers;J. F. Johnson;C. Fell
方法凝血酶效价测定。在28 ℃下，用于该分析的反应混合物由以下组合组成：牛肺促凝血酶原激酶，0.50 ml; CaCl 2（0.04 M），0.25 ml;血浆或血清等，0.50 ml试剂空间，0.25 ml。试剂空间用于在血浆评价的情况下添加生理盐水，在没有凝血酶原的标本的情况下添加纯化的凝血酶原。如果凝血酶原在反应混合物的其它试剂中可用，则该空间可用于任何所需的材料。如果使用纯化的凝血酶原溶液，其浓度必须是最终混合物中所需浓度的六倍。在所有情况下，凝血酶原的单位均表示为最终反应混合物的u/ml。为了进行分析，将0.1 ml该反应混合物加入到0.4 ml标准化纤维蛋白原溶液中，就像定量凝血酶分析一样（7）。对于结果的这种表达，以凝固时间（以秒为单位）作为纵坐标，以反应混合物中的注射分钟数作为横坐标来构建图。然后研究该曲线的凝血酶原的活化和随后的凝血酶失活的表达。去乙酰化犬血浆，乙醚处理。通过双注射器技术从健康犬中采集血液，该技术最大限度地减少了组织促凝血酶原激酶对血液的污染。作为抗凝剂，将1.85%草酸钾与7份血液混合，或将3.2%柠檬酸钠与血液以I：9的比例混合。将血液抽入含有抗凝剂的注射器中。通过以2，500 g离心15分钟获得血浆。血浆脱乙酰基如下：0.5 ml血浆+0.4 ml盐水+0.1 ml凝血酶。凝血酶质量高，含量为100 u/ml。通过机械方法去除纤维蛋白。
METHODSThrombin Titer Test. The reaction mixture, at 28OC, for this analysis consists of the following combination: bovine lung thromboplastin, 0.50 ml; CaClz (0.04 M), 0.25 ml; plasma or serum, etc., 0.50 ml reagent space, 0.25 ml.The reagent space is for the addition of physiological saline in the case of plasma evaluations and purified prothrombin in the case of specimens’ devoid of prothrombin. If prothrombin is available in other reagents of the reaction mixture this space may be utilized for any material desired. If a solution of purified prothrombin is used it must be six times as strong as the concentration desired in the final mixture. The units of prothrombin are expressed as u/ml of the final reaction mixture in all cases. For the analysis, 0.1 ml of this reaction mixture is added to 0.4 ml of standardized fibrinogen solution, just as one does for quantitative thrombin analysis (7). For this expression of the results a graph is constructed with clotting time, in seconds, as the ordinate and minutes of inmbation in the reaction mixture as the abscissa. The curve is then studied for its expression of the activation of prothrombin and the subsequent inactivation of the thrombin. Defibrinated Dog Plasma, Ether-Treated. Blood was collected from healthy dogs by means of a twosyringe technique which minimizes contamination of the blood with tissue thromboplastin. As anticoagulant, either 1.85% potassium oxalate was mixed with seven parts of blood or 3.2% sodium citrate was mixed with blood in a I: 9 ratio. The blood was drawn into the syringe containing the anticoagulant. The plasma was obtained by centrifugation at 2,500 g for 15 minutes. The plasma was defibrinated as follows: 0.5 ml plasma+ 0.4 ml saline+ 0.1 ml thrombin. The thrombin was of high quality and contained IOO u/ml. The fibrin was removed by mechanical means.