5 μg of α-syn GST fusion protein on beads were

resuspended in a buffer containing 20 mM HEPES, 1 mM ATP, 10 mM MgC₂2,

2 mM DTT, pH 7.4.

The reaction was initiated by adding 1 ug of human

recombinant PLK2 (Abcam, Cat#ab102108) and incubating at 30 °C with orbital

shaking overnight (Eppendorf ThermoMixer F1.5).

Samples were then analyzed by

NuPAGE and immunoblotted or alternatively analyzed by LC-MS/MS