Determination of ceramides and diglycerides by the diglyceride kinase assay.

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  • Author(s): Bielawska A;Bielawska A; Perry DK; Hannun YA
  • Source:
    Analytical biochemistry [Anal Biochem] 2001 Nov 15; Vol. 298 (2), pp. 141-50.
  • Publication Type:
    Comparative Study; Journal Article; Research Support, U.S. Gov't, P.H.S.; Review
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Elsevier Country of Publication: United States NLM ID: 0370535 Publication Model: Print Cited Medium: Print ISSN: 0003-2697 (Print) Linking ISSN: 00032697 NLM ISO Abbreviation: Anal Biochem Subsets: MEDLINE
    • Publication Information:
      Publication: <2000- > : San Diego, CA : Elsevier
      Original Publication: Orlando Fl : Academic Press
    • Subject Terms:
      Ceramides/*analysis ; Diacylglycerol Kinase/*analysis ; Diglycerides/*analysis; Animals ; Chromatography, High Pressure Liquid/methods ; Humans
    • Abstract:
      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.
    • Number of References:
      67
    • Grant Information:
      HL-43707 United States HL NHLBI NIH HHS; R01-GM43825 United States GM NIGMS NIH HHS
    • Accession Number:
      0 (Ceramides)
      0 (Diglycerides)
      EC 2.7.1.107 (Diacylglycerol Kinase)
    • Publication Date:
      Date Created: 20020105 Date Completed: 20020314 Latest Revision: 20071114
    • Publication Date:
      20221213
    • Accession Number:
      10.1006/abio.2001.5342
    • Accession Number:
      11757501