| Assay Method Information | |
| | Z-lyte Assay |
| Description: | GSK3B: This is a non-radioactive assay using fluorescence resonance energy transfer (FRET) between coumarin and fluorescein to detect kinase activity which is represented as ratio of coumarin emission/fluorescein emission.Briefly, recombinant GSK3B kinase, ATP and Ser/Thr peptide 09 are prepared in 1x Kinase buffer to final concentrations of 0.04 ug/mL, 46 uM, and 4 uM respectively. The mixture is allowed to incubate with the representative compounds for one hour at room temperature. All reactions were performed in duplicate. After incubation, diluted Development Buffer is added to the reaction and allowed to further incubate for one hour at room temperature. The plate is read at Ex 400 Em 455 to detect the coumarin signal and Ex 400 Em 520 to measure the signal (EnVision Multilabel Plate Reader, PerkinElmer). |
| Affinity data for this assay | |
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