Fig. 5. PIP kinase activity of the C-terminal domain of PIPkinA.GST alone or a fusion protein of GST and the C-terminal domain of PIPkinA was purified by elution from glutathione–Sepharose 4B beads, and incubated with PIP in the presence of [γ-³²P]ATP for 30 min at 22°C. Recombinant human type IIα PIP kinase was used as a control. Lipids were extracted into chloroform/methanol and the generation of radiolabelled PIP₂ was assessed by TLC.