Yeast Cell Culture and Radiolabeling of Phosphoinositides
2:58
Deacylation and Extraction of the Radiolabeled Phosphoinositides
5:14
Separation of Tritium-labeled Glycero-inositides by High Pressure Liquid Chromatography (HPLC)
7:15
Data Analysis
8:55
Results: HPLC Analysis of Radiolabeled Phosphoinositides
9:55
Conclusion
필기록
The overall goal of this procedure is to measure the amount of each phosphoinositide species in cells under various genetic and environmental conditions in order to understand phosphoinositide function and regulation. Phosphoinositide lipids contr
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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.