Tracing sphingolipid metabolism kinetics using radioactive and non-radioactive labels

Cells continuously metabolize lipids, converting them into diverse molecules that influence cellular functions- in health and disease. Sphingolipids have been specifically linked to a wide range of - cellular processes and have been shown to play a crucial role in the onset and progression of various diseases. Here, we detail a method for measuring lipid flux through a combination of labeled lipids and thin-layer chromatography (TLC). This method describes the exogenous addition of radioactive and fluorescent lipid analogues and the subsequent tracing of their metabolism. This metabolic tracing allows for conclusions to be drawn about the fate of a lipid in a given system. Though the described method focuses on sphingolipids, many alternatives are discussed for adaptation of the method to measurement of any specific lipid of interest. Overall, this method offers a robust, easy-to-adapt approach for measuring lipid flux across different cell types and experimental systems.