ADP Hunter™ HS assay is a unique tool for screening kinases and extends the range of ADP detection down to 300 nM ADP without compromising Z'. The assay employs a generic, non-antibody based, non-radioactive method to detect the amount of ADP produced as a result of enzyme activity. The amount of ADP produced is directly proportional to enzyme phoshphotransferase activity and is measured as a robust red-shifted fluorescence emission signal. The assay has a broad ATP tolerance over 2.5 logs and robust signal to background ratios, and are compatible with unmodified peptides and whole protein substrates. The assay can also be applied to detect other dNDPs such as UDP and GDP, and have been broadly applied to other types of enzymes that utilize ATP, such as ATPases.
ADP Hunter™ HS is designed for HTS and has a very low compound interference rate. With the enhanced sensitivity, the assay is ideal for running kinase assays at low kinase or ATP concentrations, or with low activity kinases. The assay has been validated in both 96- and 384-well formats.
Features and Benefits:
» Generic, non-radioactive, non-antibody based solution for all kinases
Ideal for low activity enzymes
» High Sensitivity Can run at 300 nM to 20 uM ADP
» Compatible with both peptide substrates and whole proteins
» Homogeneous and convenient
Two simple additions
Easily adapted to automation with no mixing or separation protocols
Use existing liquid handlers with a wide range of Fluorescent Intensity microplate readers