| Assay Method Information | |
| | PDK1 Enzymatic Assay |
| Description: | The experimental batches are carried out in a flashplate system with 384 wells/microtitration plate.In each case, the PDK1 sample His6-PDK1(Δ1-50)(3.4 nM) (His6 disclosed as SEQ ID NO: 2), the PDK1 substrate biotin-bA-bAKTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC (SEQ ID NO: 3) (400 nM), 4 uM ATP (with 0.2 uCi of 33P-ATP/well) and the test substance in 50 ul of conventional experimental solution per well are incubated at 30° C. for 60 min. The test substances are employed in corresponding concentrations (if desired in a dilution series). The control is carried out without test substance. The reaction is stopped using standard methods and washed. The activity of the kinase is measured via the incorporated radioactivity in top count. In order to determine the non-specific kinase reaction (blank value), the experimental batches are carried out in the presence of 100 nM staurosporine. |
| Affinity data for this assay | |
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