ab128503 PI3 Kinase Assay Kit - Abcam

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1. IntroductionPhosphoinositide 3-kinases (PI3Ks) are a family of enzymes thatplay a pivotal role in important cellular regulatory mechanisms.PI3Ks are capable of phosphorylating the 3’-OH position ofphosphoinositide lipids (PIs) generating lipid second messengers.Their function has been linked to the regulation of numerousbiological processes including cell growth, differentiation, survival,proliferation, migration. On the basis of structural similarities andsubstrate specificity, the PI3K family is divided into three classestermed I, II, and III.All human class I members are heterodimers consisting of a catalyticsubunit (MW approx. 110 kDa) and a non-catalytic subunit (MW 50,55, 85, 87 or 101 kDa). They are known to phosphorylatephosphatidylinositol (PI), phosphatidylinositol-4-mono-phosphate(PIP) and phosphatidylinositol-4,5-bisphosphate (PIP2) in vitro buthave a strong preference for PIP2 in vivo.Abcam’s PI3 Kinase Assay Kit provides a highly efficient andeasy-to-handle tool for the sensitive detection and quantification ofPI 3-Kinase activity or the screening for PI 3-Kinase inhibitors.The activity of PI3-Kinases is measured by the amount ofradioactively labeled [γ 32 P]-ATP incorporated into lipid substrate. Thesubstrate is thereafter separated using thin layer chromatography(TLC) and its radioactivity detected in a scintillation counter.3
2. Assay SummaryPrepare [γ 32 P] ATP MixPrepare Lipid Substrate VesiclesPrepare Kinase Assay MixAdd [γ 32 P] ATP MixIncubate for 15 min at 37°CStop ReactionExtract LipidsQuantify Using a Scintillation Counter4
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Page 6 and 7: 3. Components and StorageA. Kit Con
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Page 12: UK, EU and ROWEmail: technical@abca

ab128503 PI3 Kinase Assay Kit - Abcam

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