用含底物SDS-PAGE检测明胶酶活性

将明胶加入ＳＤＳ－聚丙烯酰胺凝胶中，电泳后样品中所含明胶酶按分子量大小不同停留于凝胶中的特定位置。用ＴｒｉｔｏｎＸ－１００除去ＳＤＳ使酶复性后，明胶酶便可分解凝胶中的底物，底物被分解处考马斯亮蓝染色时不着色，因此在蓝色背景下可见酶所在位置呈白色条带，根据白色条带的深浅和宽窄可判断酶活性的高低，用此方法检查了大鼠不同组织中的明胶酶活性及传代培养对血管平滑肌细胞分泌明胶酶的影响。结果显示，大鼠不同组织

Analysis of gelatinase activity by SDS-PAGE containing substrate

Gelatin the substrate of gelatinase was added to SDS-polyacrylamide gel at the time of casting After electrophoresis the gelatinases contained in samples stayed at the specified positions in the gel according to their molecular weight Then the gel was washed with TritonX-100 to remove SDS and restore the enzyme activity After reaction the substrate in the gel could be degraded The positions where the enzyme existed couldn't be stained by Coomassie brilliant blue so some clear bands appeared against the blue-stained background The activity of the enzyme could be measured by the thickness and width of the bands Gelatinase activity in various tissues of rat and different passages of VSMC was detected by this method The results suggest that gelatinase activity in kidney and aortic tissues is much higher than that in myocardium lung and liver tissues Moreover its activity in young rat is higher than that in the correspondent tissues of the old one In addition there is a type difference of gelatinase between aortic tissue and cultured VSMC