The PIP3 Mass ELISA detects PI(3,4,5)P3 (PIP3), extracted from cells, by means of a competitive 96-well ELISA assay. Assay Range: 125 to 0.04 pmol PIP3 Sample Type: Dried down lipid extracted from cells Sample Volume: 60 µL/sample, single point
Product Background The PIP3 Mass ELISA kit is a competitive ELISA for the detection of PIP3 from cell samples. Once PIP3 has been extracted from millions of cells, it is incubated with a PIP3 detector protein, before being added to a PIP3-coated microplate for competitive binding. A peroxidase-linked secondary detection reagent and colorimetric substrate is used to detect the PIP3 detector protein bound to the plate. The colorimetric signal is inversely proportional to the amount of PIP3 extracted from cells. The production of PIP3 by class I PI3-kinases (PI3-K) is important in multiple cell signaling pathways. Typically, experiments to measure PI3-K activity have involved phosphorylation of a phosphoinositide substrate using 32P, then extraction of radioactive products, and separation using thin-layer chromatography. The PIP3 Mass ELISA kit allows the user to determine PI3-K activity by measuring the amount of PIP3 extracted from cells by means of standard ELISA format, eliminating the need for radioactivity, and thin layer chromatography. Featured in Publications 1. King, JS., Teo, R., et al. The Mood Stabilizer Lithium Suppresses PIP3 Signaling in Dictyostelium and Human Cells. Disease Models and Mechanisms 2(5-6):306-12 2. Beneteau, M., Pizon, M., et al. Localization og Fas/CD95 into the Lipid Rafts on Down-Modulation of the Phosphatidylinositol 3-Kinase Signaling Pathway. Mol Cancer Res 6(4):604-613 3. Costa, C., Ebi, H., et al. Measurement of PIP3 Levels Reveals an Unexpected Role for p110beta in Early Adaptive Responses to p110alpha-Specific Inhibitors in Luminal Breast Cancer Cell 27(1):97-108 4. Gross, C., C.-W. Chang, et al. (2015). Increased Expression of the PI3K Enhancer PIKE Mediates Deficits in Synaptic Plasticity and Behavior in Fragile X Syndrome. Cell Reports, DOI:10.1016/j.celrep.2015.03.060 5. S. Choi, et al. Agonist-stimulated phosphatidylinositol-3,4,5-trisphosphate generation by scaffolded phosphoinositide kinases Nature Cell Biology (2016) 18(12), 1324-35, doi:10.1038/ncb3441 Keywords: Phosphatidylinositol (3,4,5)-trisphosphate, PtdIns(3,4,5)P3, PI(3,4,5)P3, PIP3, Phosphatidylinositol 3-kinase, PI 3-kinase, PI3-Kinase, PI3-K, PI3K, PI3 Kinase, PI3 K, cell extraction, lipid extraction
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