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17-495 | PIP3 Quantification TR-FRET Assay

1 plate  Kit capacity: 96-wells
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      Key Specifications Table

      Key ApplicationsAnalytes AvailableDetection Methods
      KA PI 3-Kinase TR-Fret, HTRF
      Catalogue Number17-495
      DescriptionPIP3 Quantification TR-FRET Assay
      OverviewPI 3-Kinases α, β, γ and δ (h) catalyse the phosphorylation of phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidylinositol 3,4,5-trisphosphate (PIP3) in the presence of ATP and Mg2+. The PIP3 product is detected by displacement of biotin-PIP3 from an energy transfer complex consisting of Europium labelled anti-GST monoclonal antibody, a GSTtagged pleckstrin homology (PH) domain, biotinylated PIP3 and Streptavidin-Allophycocyanin (APC). Excitation of Europium in the complex results in an energy transfer to the APC and a fluorescent emission at 665 nm. The PIP3 product isolated from cell membranes displaces biotin-PIP3 from the complex resulting in a loss of energy transfer and thus a decrease in signal.
      Product Information
      • Black 96-well plate
      • Detection Buffer
      • DM B
      • DM A
      • Stop B
      • PIP3 Standard
      • Lipid Recovery Buffer
      • 1M DTT
      Detection methodTR-Fret HTRF
      ApplicationThe PIP3 Quantification TR-FRET Assay kit provides an easy to use method for assaying PIP3 levels by detecting the displacement of biotin-PIP3 from an energy transfer complex.
      Key Applications
      • Kinase Assay
      Application NotesThis assay requires lipid extraction from cells. A protocol is included with this assay.
      Biological Information
      Analytes Available
      • PI 3-Kinase
      Entrez Gene Number
      Entrez Gene SummaryPhosphatidylinositol 3-kinase is composed of an 85 kDa regulatory subunit and a 110 kDa catalytic subunit. The protein encoded by this gene represents the catalytic subunit, which uses ATP to phosphorylate PtdIns, PtdIns4P and PtdIns(4,5)P2. This gene has been found to be oncogenic and has been implicated in cervical cancers.
      Gene Symbol
      • p110-alpha
      • MGC142161
      • PI3K
      • MGC142163
      • PIK3CA
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P42336 # Phosphorylates PtdIns, PtdIns4P and PtdIns(4,5)P2 with a preference for PtdIns(4,5)P2.
      SIZE: 1068 amino acids; 124284 Da
      SUBUNIT: Heterodimer of a p110 (catalytic) and a p85 (regulatory) subunit. Binds to IRS1 in nuclear extracts. Interacts with RUFY3 (By similarity).
      DISEASE: SwissProt: P42336 # Defects in PIK3CA are associated with colorectal cancer (CRC) [MIM:114500]. & Defects in PIK3CA are associated with breast cancer [MIM:114480]. & Defects in PIK3CA are associated with epithelial ovarian cancer (EOC) [MIM:604370].
      SIMILARITY: SwissProt: P42336 ## Belongs to the PI3/PI4-kinase family. & Contains 1 C2 domain. & Contains 1 PI3K/PI4K domain.
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Packaging Information
      Material Size1 plate
      Material PackageKit capacity: 96-wells
      Transport Information
      Supplemental Information

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      Life Science Research > Protein Detection and Quantification > Activity Assays > Kinase Assays