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Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation
Chapters
- 00:05Title
- 00:58Yeast Cell Culture and Radiolabeling of Phosphoinositides
- 02:58Deacylation and Extraction of the Radiolabeled Phosphoinositides
- 05:14Separation of Tritium-labeled Glycero-inositides by High Pressure Liquid Chromatography (HPLC)
- 07:15Data Analysis
- 08:55Results: HPLC Analysis of Radiolabeled Phosphoinositides
- 09:55Conclusion
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.
