Enzymatic fluorometric assay for myo-inositol trisphosphate

James A. Shayman, Aubrey R. Morrison, Oliver H. Lowry

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

The determination of myo-inositol trisphosphate by an enzymatic fluorometric assay is presented. The method involves the acid extraction of water-soluble inositol polyphosphates followed by separation by anion-exchange chromatography. Samples are subsequently neutralized by passage over a Dowex Cl- resin and elution with lithium chloride. Samples are then desalted with ethanol. Following dephosphorylation with alkaline phosphatase, free myo-inositol is measured enzymatically via the NAD-dependent oxidation to scyllo-inosose with myo-inositol dehydrogenase. The efficiency of recovery, assay specificity, and an application to the measurement of inositol polyphosphates in hormone-stimulated tissue are discussed.

Original languageEnglish
Pages (from-to)562-568
Number of pages7
JournalAnalytical Biochemistry
Volume162
Issue number2
DOIs
StatePublished - May 1 1987

Keywords

  • enzymatic assay
  • fluorometry
  • inositol polyphosphate

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