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 language | English |
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Pages (from-to) | 562-568 |
Number of pages | 7 |
Journal | Analytical Biochemistry |
Volume | 162 |
Issue number | 2 |
DOIs | |
State | Published - May 1 1987 |
Keywords
- enzymatic assay
- fluorometry
- inositol polyphosphate