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Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
 

Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation

Article DOI: 10.3791/53529-v • 10:52 min • January 6th, 2016
January 6th, 2016 •
*1, *1, 1
1Department of Chemistry and Biology, Program in Molecular Science, Ryerson University
* These authors contributed equally

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Phosphoinositides are signaling lipids whose relative abundance rapidly changes in response to various stimuli. This article describes a method to measure the abundance of phosphoinositides by metabolically labeling cells with 3H-myo-inositol, followed by extraction and deacylation. Extracted glycero-inositides are then separated by high-performance liquid chromatography and quantified by flow scintillation.

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Keywords: Radiolabeling Phosphoinositides HPLC Flow Scintillation Yeast Mammalian Cells Inositol Perchloric Acid EDTA Centrifugation Sonication Liquid Culture Cell Growth Cell Lysis
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