Phosphatidylinositol 3-kinase alpha E545K mutant and Regulatory Subunit 1
U79143 (PI3 kinase alpha) - NM_181523 (p85)
PI3K alpha E545k + P85 (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 80 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.
PI3kinasealpha_E545K-p85.pdf (92.84 KB)