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Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H]Inositol Labeling Protocols in Permeabilized Cells

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Calcium Signaling Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 937))

Abstract

Data on the production of inositol phosphates is a useful complement to measurements of intracellular Ca2+. The basic principle is labeling of the inositol lipids by growing the appropriate cell line in culture in the presence of [3H]inositol for 2–3 days to reach labeling equilibrium. Lithium ions at 10 mM inhibits the degradation of inositol phosphates to free inositol and is used to trap the inositol in the inositol polyphosphate forms. Inositol phosphates can be separated with ease from free inositol by using anion exchange chromatography. A method capable of easily processing approximately 40–60 samples in a single day is presented.

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Authors and Affiliations

  1. Department of Neuroscience, Physiology, and Pharmacology, University College London, London, UK

    Alison Skippen, Philip Swigart & Shamshad Cockcroft

Authors
  1. Alison Skippen
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  2. Philip Swigart
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  3. Shamshad Cockcroft
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Corresponding author

Correspondence to Shamshad Cockcroft .

Editor information

Editors and Affiliations

  1. , Department of Cardiovascular Sciences, Leicester Royal Infirmary, Infirmary Square, Leicester, LE1 5WW, Leicestershire, United Kingdom

    David G. Lambert

  2. , Department of Cardiovascular Sciences, Leicester Royal Infirmary, Infirmary Square, Leicester, LE1 5WW, Leicestershire, United Kingdom

    Richard D. Rainbow

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Skippen, A., Swigart, P., Cockcroft, S. (2013). Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H]Inositol Labeling Protocols in Permeabilized Cells. In: Lambert, D., Rainbow, R. (eds) Calcium Signaling Protocols. Methods in Molecular Biology, vol 937. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-086-1_9

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  • DOI: https://doi.org/10.1007/978-1-62703-086-1_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-085-4

  • Online ISBN: 978-1-62703-086-1

  • eBook Packages: Springer Protocols

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