Determination of ceramides and diglycerides by the diglyceride kinase assay.
This review will discuss the utilization of the diglyceride (DG) kinase assay as an analytical method that allows the simultaneous quantitation of DG and ceramide from cell and tissue samples. This enzymatic approach is sensitive, quantitative, and linear over a broad range (20 pmol to 20-25 nmol) for both classes of lipids. It is also practical in that it can be applied to crude lipid extracts and used to process many samples (up to 100) concurrently. However, it has become apparent that this assay has not been conducted optimally, primarily as a result of lack of adherence to its basic principles. The principles illustrated here are also useful to all enzymatic quantitative methods.
Bielawska
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Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston 29425, USA. bielawsk@musc.edu
Perry
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DK
Hannun
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YA
D000818:Animals; D002518:Ceramides / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D019852:Diacylglycerol Kinase / Q000032:analysis*; D004075:Diglycerides / Q000032:analysis*; D006801:Humans
D002518:Ceramides; D004075:Diglycerides; D019852:Diacylglycerol Kinase