| Assay Method Information | |
| | Protein Crystallography Inhibitory Activity Assay |
| Description: | Aurora A was exchanged into 50 mM phosphate buffer (pH 7.4) including 1 mM DTT via PD-10 columns and was concentrated to 20 mg mL-1 using Amicon Ultra-4 10K centrifugal devices (Millipore; Billerica, Mass.). Aurora A crystals were grown using sitting drop vapor diffusion at 18° C. from a 1:1 volume ratio of Aurora A and reservoir solution (200 mM sodium tartrate/20% polyethylene glycol 3350 for the DFG-in inhibitors or 10% Tacsimate/20% polyethylene glycol 3350 for the DFG-out inhibitors) with a final inhibitor concentration of 1 mM. Crystals appeared after two days and were allowed to grow for another 2 days. For data collection, crystals were harvested in a cryoprotectant mixture consisting of the respective reservoir composition including 1 mM inhibitor, 50 mM phosphate pH (7.4) and 25% (v/v) ethylene glycol. |
| Affinity data for this assay | |
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