Fluorescent Lipids as Probes for Sphingosine Kinase Activity by Capillary Electrophoresis

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.