title: Measurement of phospholipase C by monitoring inositol phosphates using [³H]inositol labeling protocols in permeabilized cells.
abstract: Data on the production of inositol phosphates is a useful complement to measurements of intracellular Ca(2+). 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.
authors.affiliation: Department of Neuroscience, Physiology, and Pharmacology, University College London, London, UK.
authors.forename: Alison
authors.initials: A
authors.lastname: Skippen
authors.forename: Philip
authors.initials: P
authors.lastname: Swigart
authors.forename: Shamshad
authors.initials: S
authors.lastname: Cockcroft
mesh_terms: D018922:HL-60 Cells; D006801:Humans; D007294:Inositol / Q000737:chemistry* / Q000378:metabolism*; D007295:Inositol Phosphates / Q000737:chemistry / Q000378:metabolism*; D014316:Tritium / Q000737:chemistry / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: D007295:Inositol Phosphates; D014316:Tritium; D007294:Inositol; D010738:Type C Phospholipases