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Assay Method Information |
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| | Dissociation Kinetics Assay |
| Description: | A stock solution was prepared containing complex of Aurora A and Compound I (final total concentrations of 6nM and 5nM, respectively). Separately, several aliquots of MLN8237 (1.25uM) were prepared (20uL each). All mixtures were pre-incubated at 30 C for 15 min. Subsequently, 5uL of stock solution of complex was transferred into the first aliquote of MLN8237 and incubated further at 30 C; this was repeated at different time-points. After 4 hours, all the resulting mixtures were transferred onto a microtiter plate and fluorescence anisotropy was measured. The same experiment was repeated for complex consisting of Compound II and Aurora A. In the case of complex of Compound III with Aurora A, VX-689 was used as the competitive binder. |
| Affinity data for this assay | |
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