ADP-Glo kinase assay

100 nM purified ULK1 complex was mixed with 5 μM ULKtide (SignalChem Biotech Inc), and incubated at room temperature for 1 hr. The reaction buffer was 20 mM HEPES pH 8.0, 200 mM NaCl, 2 mM MgCl₂, 100 μM ATP, 20 mM Maltose and 1 mM TCEP. The reaction was terminated by adding an ATP-depletion reagent. Then a kinase detection reagent was added to convert ADP to ATP, which is used in a coupled luciferase reaction. The luminescent output was measured with a GloMax-Multi detection system (Promega) and was correlated with the kinase activity.