| Assay Method Information | |
| | Fluorescence Polarization Assay |
| Description: | Fluorescence polarization assays used GST-tagged enzyme and an ATP-competitive Rhodamine-green labelled fluoroligand. These components were dissolved in a buffer. Thirty microliters of this mixture were added to wells containing test compound and incubated for 30-60 min at room temperature. Fluorescence anisotropy was read in a Molecular Devices Acquest (excitation 485 nm/emission 535 nm). All values quoted are means of IC50 results. Ki values were calculated from IC50s using a modified Cheng-Prusoff equation. |
| Affinity data for this assay | |
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