An assay to evaluate the capacity of cholesterol acceptors using BODIPY-cholesterol in cells.

Cholesterol is a structural component of cell membranes. Most cells are incapable of its catabolism, and intracellular cholesterol accumulation is linked to several disorders including cardiovascular and neurodegenerative diseases. Cholesterol efflux, essential to its metabolism, is dependent on acceptors such as apolipoproteins. Here, we describe an assay to evaluate the capacity of cholesterol acceptors. Cells are treated with an analog of cholesterol tagged with fluorescent BODIPY. Addition of an acceptor leads to BODIPY-cholesterol efflux, measured using a plate reader. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021).1.