The PI(4)P 5-Kinase (PIP5K1) 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 PIP5K1 activity 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 I PI(4)P 5-Kinase (PIP5KI) catalyzes the phosphorylation of phosphatidylinositol-4-phosphate [PI(4)P] to phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. PIP5KI has been shown to play a role in several cellular functions including gene transcription, actin cytoskeleton reorganization, secreation and endocytosis. It is thought that PIP5KI activity is responsible for the majority of the PI(4,5)P2 found in the cell which is an important second messenger for many cellular processes.
The assay reaction is prepared by the addition of PIP5KI (enzyme), PI(4)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(4)P 5-Kinase Activity Assay is tested using SignalChem’s PIP5KIA (P16-10AG) kinase.