title: Fluorescent lipids as probes for sphingosine kinase activity by capillary electrophoresis.
abstract: Capillary electrophoresis (CE) is one among a number of highly sensitive chemical separation techniques used to characterize single or a small number of cells and to develop assays of enzymatic activity. Other commonly used techniques include mass spectrometry and electrochemistry; however, CE using laser-induced fluorescence detection (LIF) is the most sensitive of these techniques. In CE-LIF, fluorescently labeled proteins or lipids are normally separated based on their size to charge ratio in the interior of a small capillary filled with an electrolyte upon the application of an electric field. In this chapter, we describe the application of CE-LIF for the determination of the bioactivity of fluorescently lipids and sphingosine kinase activity.
authors: Department of Chemistry, Kent State University, Kent, OH, USA.
authors: Philip M
authors: PM
authors: Yangyuoru
authors: Latanya
authors: L
authors: Hammonds-Odie
authors: Simon M
authors: SM
authors: Mwongela
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods; D057075:Enzyme Assays / Q000379:methods; D019793:Fluorescein / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D008055:Lipids / Q000737:chemistry*; D008246:Lysophospholipids / Q000737:chemistry; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013110:Sphingosine / Q000031:analogs & derivatives / Q000737:chemistry; D000099157:Sphingosine Kinase
chemical_list: D019793:Fluorescein; D005456:Fluorescent Dyes; D008055:Lipids; D008246:Lysophospholipids; D017853:Phosphotransferases (Alcohol Group Acceptor); D013110:Sphingosine; D000099157:Sphingosine Kinase; C060506:sphingosine 1-phosphate