Protocol to determine in vitro lipid-protein interactions using liposome flotation assay

Lipid-protein interactions are crucial for cellular metabolism, transport, and signaling. Here, we present a detailed protocol to determine lipid-protein interactions using purified proteins, commercially available lipids, and a gradient centrifugation-based technique. We show how to make liposomes, test liposome size, form sucrose gradients with liposomes and purified protein, and collect and analyze the fractions after centrifugation. This protocol can determine lipid-protein interactions using a variety of liposome compositions for different proteins. For complete details on the use and execution of this protocol, please refer to House et al.¹.