| Assay Method Information | |
| | LATS1 High ATP HTRF Assay |
| Description: | The LATS1 biochemical assay was performed using the HTRF KinEASE-STK S1 Kit (Cisbio, Cat #62ST1PEC), following the protocol from the manufacturer. Compounds were dispensed by the Echo Liquid Handler (Labcyte) into a white 384-well plate (PerkinElmer, Cat #6008289). 3 uL of 2xLATS1 enzyme solution was added to the compounds, followed by a 10 minute incubation at room temperature. Then, 3 uL of 2xATP and STK S1 peptide solution was added to initiate the enzyme reaction for one hour at room temperature. The final condition of the reaction was 0.025 nM LATS1, 5000 uM ATP, 0.5 uM STK S1 peptide in 50 mM HEPES pH 7.2, 10 mM MgCl2, 0.1% BGG, 0.005% Brij-35, 1 mM DTT. The reaction was quenched and the HTRF signal was read. |
| Affinity data for this assay | |
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