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.|
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.