A highly sensitive chromogenic microtiter plate assay for plasminogen activators which quantitatively discriminates between the urokinase and tissue-type activators.
A highly sensitive chromogenic microtiter plate assay for plasminogen activators which quantitatively discriminates between the urokinase and tissue-type activators.
复制标题
用于纤溶酶原激活剂的高灵敏度显色微量滴定板测定，可定量区分尿激酶和组织型激活剂。
DOI：
10.1016/0006-291x(87)90463-3
复制
发表时间：
1987
影响因子：
3.1
通讯作者：
Littlefield,BA
中科院分区：
文献类型：
--
作者：
Karlan,BY;Clark,AS;Littlefield,BA
描述了一种简单、高灵敏度的纤溶酶原激活剂显色微滴板测定方法。该实验是基于纤溶蛋白切割合成的三肽纤溶蛋白底物h - d -去甲酰基-六氢酪氨酸对硝基苯胺，产生黄色发色团对硝基苯胺。后一种化合物的产生，然后在酶联免疫吸附测定仪上分光光度测定405 nm。在适当的孵育时间下，可在单次实验（0.01-100 milliPloug单位）中实现至少四个数量级的线性测定。利用组织型纤溶酶原激活剂对纤维蛋白的酶活性依赖性，选择性使用可溶性纤维蛋白产品可以区分尿激酶和组织型激活剂。这方面的分析方法用于分析含有两种类型纤溶酶原活性物的复杂样品。
A simple and highly sensitive chromogenic microtiter plate assay for plasminogen activators is described. The assay is based on plasmin cleavage of the synthetic tripeptide plasmin substrate H-D-norleucyl-hexahydrotyrosyllysine p-nitroaniline, which yields the yellow chromophore p-nitroanilide. Production of the latter compound is then quantitated spectrophotometrically at 405 nm on an ELISA plate reader. Linearity of the assay can be achieved over at least four orders of magnitude in a single experiment (0.01–100 milliPloug units) with appropriate incubation times. Capitalizing on tissue-type plasminogen activator's dependence on fibrin for enzymatic activity, the selective use of soluble fibrin products allows discrimination between urokinase and tissue-type activator. The utility of this aspect of the assay for the analysis of complex samples containing both types of plasminogen activitors is demonstrated.