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
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.
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