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Enzymatic fluorometric assay for myo-inositol trisphosphate

✍ Scribed by James A. Shayman; Aubrey R. Morrison; Oliver H. Lowry

Publisher: Elsevier Science
Year: 1987
Tongue: English
Weight: 578 KB
Volume: 162
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(87)90434-9

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✦ Synopsis

The determination of my@inositol t&phosphate 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 c1-resin and elution with lithium chloride. Samples are then desalted with ethanol. Following dephosphorylation with alkaline phosphatase, free my&nosi-to1 is measured enxymatically via the NAD-dependent oxidation to scy1/&nosose with myoinositol dehydrogenase. The efficiency of recovery, assay specificity, and an application to the measurement of inositol polyphosphates in hormone-stimulated tissue are discussed. 8 1987 Academic F'ms, Inc.

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