The PI(5)P 4-Kinase (PIP4KII) Activity Assay is an ATP depletion assay which quantifies the remaining ATP levels in solution following the kinase reaction. The assay can be used to quantify PI(5)P 4-Kinase in vitro or in cell lysate.
Sample Volume: 10 µL of the kinase samples per well
Assay Incubation Time: 30 min to 2 hrs depending on enzyme reaction time
Assay Range: 0.15 µM to 10 µM ATP
Type II PI(5)P 4-Kinase (PIP4KII) catalyzes the phosphorylation of phosphatidylinositol-5-phosphate [PI(5)P] to phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. PIP4KII has been shown to play a role in certain types of breast and colon cancer in addition to being important in several physiological processes such as insulin signaling, stress response, and embryonic development in several model organisms. It has also been implicated as an important regulator of nuclear PI(5)P levels.
The assay reaction is prepared by the addition of PIP4KII (enzyme), PI(5)P (substrate) in reaction buffer and initiated by the addition of ATP. The reaction is then stopped by adding the ATP detector after the chosen reaction time. The luminescent signal is inversely correlated with the kinase activity. Echelon’s PI(5)P 4-Kinase (PIP4KII) Activity Assay is tested using SignalChem’s PIP4KIIA (P76-10AG) kinase.