2x Reaction Buffer (2 x 1.5 ml)
1mM ATP (400 l)
Lipid Substrate (8 tubes)
Control PI 3-Kinase (5 g) Description
PI 3-Kinase Assay Kit provides a highly efﬁcient and easy-to-handle tool for the sensitive detection and quantiﬁcation of PI 3-Kinase activity or the screening for PI 3-Kinase inhibitors. The activity of PI3-Kinases is measured by the amount of radioactively labeled [32P]-ATP incorporated into lipid substrate. The substrate is thereafter separated using thin layer chromatography (TLC) and its radioactivity detected in a scintillation counter. Background
Phosphoinositide 3-kinases (PI3Ks) are a family of enzymes that play a pivotal role in important cellular regulatory mechanisms. PI3Ks are capable of phosphorylating the 3’-OH position of phosphoinositide lipids (PIs) generating lipid second messengers. Their function has been linked to the regulation of numerous biological processes including cell growth, differentiation, survival,
proliferation, migration. On the basis of structural similarities and substrate speciﬁcity, the PI3K family is divided into three classes termed I, II, and III. All human class I members are heterodimers consisting of a catalytic subunit (MW approx. 110 kDa) and a
non-catalytic subunit (MW 50, 55, 85, 87 or 101 kDa). They are known to phosphorylate phosphatidylinositol (PI), phosphatidylinositol-4-mono-phosphate (PIP) and phosphatidylinositol-4,5-bisphosphate (PIP2) in vitro but have a strong preference for PIP2 in vivo.