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Transferases, and especially kinases, are one of the most important and extensively studied enzyme families in biochemical and medical research due to their physiological relevance, diversity, and ubiquity. Many currently available kinase assays work well with enzymes that consume large amounts of ATP during the reaction, but are not sensitive enough to detect small changes in the amount of ATP, which limits the detection of kinases with low ATP turnover rates, such as growth factor receptor kinases, thus requiring a highly sensitive assay to screen for low-activity kinases.

Overview :

The Rhinogen ^® Kinase-Turbo™ ADP Kinase Activity Assay Kit is a luminescent ADP assay kit that provides a universal, homogeneous, high-throughput screening method to measure kinase activity by quantifying the amount of ADP produced during the kinase reaction. The Rhinogen ^® Kinase-Turbo™ ADP Kinase Activity Assay Kit can be used to monitor the activity of nearly any kinase that can produce ADP. The Rhinogen ^® Kinase-Turbo™ ADP Kinase Activity Assay Kit measures the activity of enzymes such as kinases or ATPases that use 0-10μM ATP. The ADP Kinase assay is performed in multi-well plates, allowing the detection of kinase activity in reaction volumes as low as 5μl.

Figure 1. Principle of Kinase-Turbo™ ADP Kinase Activity Assay Kit

The assay is performed in two steps :
1) First, after the kinase reaction, an equal volume of the First reagent is added to terminate the kinase reaction and deplete the remaining ATP. 2) Second, the Second reagent is added to simultaneously convert ADP to ATP and measure the newly generated ATP using the luciferase/luciferin reaction. The resulting luminescent signal is measured using a microplate reader. The luminescent value can be correlated to the ADP concentration using the ATP to ADP conversion curve.