酶显色底物ADP-ribose-pNP

酶显色底物ADP-ribose-pNP是美国AAT Bioquest生产的用于过氧化物酶的试剂，ADP-核糖-pNP是用来评估多聚ADP-核糖聚合酶（PARP）活性的一种显色底物。在405nm处可检测释放的p-硝基酚吸光度。标准曲线的斜率通常用来转换产生的ADP-核糖-pNP比色底物的的吸光度。使用ADP-核糖-pNP作为显色底物，PARP-1具有大Km和Vmax数值（分别为151 uM和1.30nmolmin-1mg-1），其次是Tankyrase-1(82uM和18pmolmin-1mg-l)和VPARP（46uM和2 pmolmin-1mg-l）。显色底物通常用来决定PARP-1,tankyrase-1和VPARP 的动力学参数，然后筛选PARP-1,tankyrase-1和VPARP小分子抑制剂。由于ADP-核糖-pNP连续实验分析比标准的非连续PARP试验更有优势，因此其可用于高通量筛选PARP-1,tankyrase-1和VPARP的活性以及相关抑制剂。百萤生物是AAT Bioquest的中国代理商，为您提供优质的酶显色底物ADP-ribose-pNP。 

适用仪器

样品实验方案 

溶液制备 

1.储备溶液

所有未使用的储备溶液应分成一次性等分试样，并在制备后储存在-20°C。 避免反复冻融循环。
1.1 ADP-核糖-pNP原液：
在H₂O中制备5-10mM的储备溶液。 注意：应立即使用原液。

2.工作溶液

ADP-核糖-pNP工作溶液：
通过用测定缓冲液（50mM Tris，10mM MgCl 2，pH 8.0）稀释储备溶液来制备0.25mM测定溶液。

样品操作步骤

1.将0.01mL /孔的样品溶液加入0.09mL /孔测定溶液中，使96孔透明板中的终体积为0.1mL。

2.使用吸光度酶标仪在405nm处监测平板。

参考文献

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