ADP-Glo? Max Assay

• 产品编号：V7002      品牌：promega       原厂货号：V7002
• 产品分类：蛋白质检测与分析 > 酶检测和活性分析 > 激酶检测
• 应用分类：

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描述：

原厂资料：

The ADP-Glo™ Max Assay is a luminescent ADP detection assay that provides a universal, homogeneous, high-throughput screening method to measure ATPase or kinase activity by quantifying the amount of ADP produced in a reaction. The assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) when higher ATP concentration is required (up to 5mM). The ADP-Glo™ Max Assay produces a strong signal that positively correlates with enzyme activity and can be adapted to a multitude of plate formats.

The assay is performed in two steps: first, after the completion of the ADP-producing reaction, an equal volume of ADP-Glo™ Reagent is added to terminate the reaction and deplete the remaining ATP. Second, the ADP-Glo™ Max Detection Reagent is added to simultaneously convert ADP to ATP, and the latter is converted to light in a coupled reaction with luciferase/luciferin.

The ADP-Glo™ Max Assay has a high dynamic range and produces a strong signal at low ATP to ADP conversion, making it well suited for screening low-activity ATPases such as drug membrane transporters and heat shock proteins. The assay produces minimal false hits and Z´ values of greater than 0.7.

Several Kinase Enzyme Systems are available. Visit www.promega.com/kinase/ to see the collection.

Features - Benefits

High Signal Strength at Low ATP Conversion: Users can measure enzyme activity that more closely mimics physiological conditions. This makes the assay very well suited for low-activity ATPases/kinases.
Sensitive: The assay is sensitive to low concentrations of ADP, thus requiring less enzyme than other assays; cost savings.
Universal: The assay can be used with virtually with any ADP-producing enzyme—enables researchers to screen a wider range of enzymes using a single platform.
Accommodate Wide Range of ATP Levels: The assay can be used at ATP concentrations up to 5mM, important for enzymes with high Km values for ATP and for mode of action studies.
Accurate: Accurately measures ADP levels at a wide range of starting ATP concentrations; users assured that activity measured truly reflects enzyme activity and produces accurate IC50s comparable to radioactivity-based assays.

Applications

Screen library compounds for effects on target enzymes.
Perform mode of action studies on enzymes of interest.
Measure activity of immunoprecipitated ATPases/kinase.

ADP-Glo™ Applications Database.

注意事项：

将整个系统储存于–20°C 。使用前，将所有组分在室温完全融化，接着充分混合后备用。由于冻融循环后，ATP 易于水解，因此应将其分装成单次使用的小份，储存于 –20°C 。将 ADP-Glo™ Max 检测试剂（ ADP-Glo™ Max 检测缓冲液 + 底物）制备好后，分装成小份，储存于–20°C。融化后， ADP-Glo™ Max 检测缓冲液可能形成沉淀，请查看技术手册的3.A 部分，了解溶解沉淀的操作方法。为方便起见，可将 ADP-Glo™ 试剂和ADP-Glo™ Max 检测试剂置于室温（22°C）24个小时，而不会损失信号。