Phosphatidylinositol 3-kinase type 1 gamma
PI3K gamma (diluted in 12.5mM Glycine-NaOH (pH 8.5), 50mM KCl, 2.5mM MgCl2, 1mM DTT, 0.05% CHAPS) is assayed in total volume of 20ul containing 12.5mM Glycine-NaOH (pH 8.5), 50mM KCl, 2.5mM MgCl2, 1mM DTT, 0.05% CHAPS, 0.01mM ATPand 0.05mM diC8 PIP2. The enzyme is assayed for 75 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 assys are incubated for 40 min and then read on PerkinElmer Envision for 1sec/well.
PI3_kinase_gamma_screening.pdf (89.12 KB)