ELECTROPHORETIC ANALYSIS OF PLASMINOGEN ACTIVATORS IN POLYACRYLAMIDE GELS CONTAINING SODIUM DODECYL-SULFATE AND COPOLYMERIZED SUBSTRATES
ELECTROPHORETIC ANALYSIS OF PLASMINOGEN ACTIVATORS IN POLYACRYLAMIDE GELS CONTAINING SODIUM DODECYL-SULFATE AND COPOLYMERIZED SUBSTRATES
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DOI：
10.1016/0003-2697(80)90338-3
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发表时间：
1980-01-01
影响因子：
2.9
通讯作者：
DOWDLE, EB
中科院分区：
文献类型：
--
作者：
HEUSSEN, C;DOWDLE, EB
本文介绍了一种新的电泳分析技术，该技术可用于在含有共聚纤溶酶原和明胶的十二烷基硫酸钠-聚丙烯酰胺凝胶中电泳分析[人]纤溶酶原激活剂。该方法依赖于这样一个事实，即酶原和明胶，当纳入到聚丙烯酰胺基质在铸造时，被保留在随后的电泳酶样品，并作为令人满意的顺序，在原位底物的纤溶酶原激活剂带的本地化负染色。使用非离子去污剂Triton X-100去除十二烷基硫酸钠并恢复酶活性。该方法可用于检测低至1 mU [单位]的尿激酶，并有效区分黑色素瘤和尿激酶型纤溶酶原激活剂。通过从凝胶中省略纤溶酶原来检测纤溶酶原非依赖性蛋白酶。
A new technique was described for the electrophoretic analysis of [human] plasminogen activators in sodium dodecyl sulfate-polyacrylamide gels containing copolymerized plasminogen and gelatin. The method depends upon the fact that the zymogen and gelatin, when incorporated into the polyacrylamide matrix at the time of casting, are retained during subsequent electrophoresis of enzyme samples and serve as satisfactory sequential, in situ substrates for the localization of plasminogen activator bands by negative staining. The nonionic detergent, Triton X-100, is used to remove sodium dodecyl sulfate and restore enzyme activity. The method can be used to detect as little as 1 mU [unit] of urokinase and effectively distinguishes between melanoma- and urokinase-type plasminogen activators. Plasminogen-independent proteases are detected by omission of plasminogen from the gel.