Phosphatidylinositol-5-phosphate 4-kinase type-2 alpha
PIP5K2A ( diluted in 12.5mM Glycine-NaOH (pH 8.5), 50mM KCl, 2.5mM MgCl2, 1mM DTT, 0.25% Na-Cholate and 1mg/ml BSA) is assayed in total volume of 20ul containing 12.5mM Glycine-NaOH (pH 8.5), 50mM KCl, 2.5mM MgCl2, 1mM DTT, 0.25% Na-Cholate, 0.01mM ATPand 0.05mM diC8 PI(5)P. The enzyme is assayed for 15 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.
PIP5K2A_5.pdf (84.69 KB)