Comparison of the luminescent ADP-Glo assay to a standard radiometric assay for measurement of protein kinase activity.
Many assay technologies have been developed and utilized to efficiently assay and screen against protein kinase targets. The radiometric assay format for assaying the protein kinase targets has been considered the "Gold Standard" format since it allows the direct readout of kinase functional activity and is a universal assay that is highly sensitive. However, the hazardous nature of the radiometric assay together with the regulatory hurdles has led to the development of alternative assay formats for assessing protein kinase activity measurements. The luminescent ADP-Glo assay has been developed as an alternative to radiometric format for assaying protein kinase targets. This assay allows the measurement of the ADP product formed during the kinase reaction. Therefore, the luminescent ADP-Glo assay is similar to the radiometric format in that it measures the direct product of the protein kinase reaction. Furthermore, since the ADP product is generated by all protein kinase reactions, this is a universal format that can be used for assaying any given protein kinase target. Analysis of data generated with multiple protein kinase targets and the luminescent ADP-Glo technology shows comparable results to the radiometric assay format. Therefore, the luminescent ADP-Glo assay is a robust new technology for evaluating catalytic function of protein kinases as well as other ATPases.
Sanghera
Jasbinder
J
SignalChem Inc., Richmond, British Columbia, Canada. jsanghera@signalchem.com
Li
Rick
R
Yan
Jun
J
D000244:Adenosine Diphosphate / Q000032:analysis* / Q000737:chemistry*; D002623:Chemistry Techniques, Analytical; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000032:analysis*; D010770:Phosphotransferases / Q000032:analysis* / Q000737:chemistry*; D025941:Protein Interaction Mapping / Q000379:methods*; D011869:Radioligand Assay / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D008164:Luminescent Proteins; D000244:Adenosine Diphosphate; D010770:Phosphotransferases