ADP-Glo™ Max Assay Technical Manual

Literature # TM343

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 ADP-Glo™ Max 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 is performed in a multiwell plate and can detect kinase or ATPase activity in a reaction volume as low as 5µl. 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 using a luciferase/luciferin reaction. Luminescence is read using a luminometer and is correlated to ADP concentrations by using an ATP-to-ADP conversion curve.