四唑蓝比色法测定肝素酶活性

Analysis of heparanase activity by colorimetry

目的:建立一种检测肝素酶活性的简便快捷的方法。方法:肝素酶作用于肝素/硫酸肝素特定位点的β-1,4糖苷键生成还原性醛基末端,还原性醛基末端可将生色底物四唑蓝还原成蓝色产物。颜色变化反映还原性醛基基团数量的改变的特性,被用于肝素酶活性的检测。结果:四唑蓝比色法的结果表明,颜色的改变程度与酶活性成正比;凝胶和SDS-PAGE电泳法验证了四唑蓝比色法的结果。结论:四唑蓝比色法可较准确地反映肝素酶活性,可以成为测定肝素酶活性的简便、经济、有效的方法。

Objective： To establish a method that can be used for the determination of heparanase activity. Methods： Aldehyde group produced by heparanase reacts with chromogenic substrate tetrazolium blue chloride, resulting in color change, which can be used to evaluate heparanase activity. Results：The change of absorbance was proportional to heparanase activity and confirmed by gel electrophoresis. Conclusion：The colorimetry as a kind of heparanase activity detection method can be used to determine heparanase activity exactly.