Diacylglycerol kinase zeta
DGK zeta (diluted in 20mM Tris- HCl, 67mM KCl, 10mM MgCl2, 1mM DTT, 0.05mg/ml BSA) is assayed in total volume of 20ul containing 20mM Tris- HCl, 67mM KCl, 10mM MgCl2, 1mM DTT, 0.05mg/ml BSA, 0.01mM ATPand 0.1mM diacyl-glycerol. The enzyme is assayed for 40 min after which 20ul of ADP-Glo reagent is added. After a further incubation of 40 min, 40ul of Kinase Detection Buffer is added. The assays are incubated for 40 min and then read on PerkinElmer Envision for 1sec/well.
DGK_zeta_screening.pdf (85.67 KB)