Separation of actin-dependent and actin-independent lipid rafts

Abstract Lipid rafts have been isolated on the basis of their resistance to various detergents and more recently by using detergent-free procedures. The actin cytoskeleton is now recognized as a dynamic regulator of lipid raft stability. We carefully analyzed the effects of the cortical actin-disrupting agent latrunculin B on lipid raft markers of both protein and lipid nature and show that two detergent-free membrane subtypes can be isolated and separated from each other on a one-step density gradient combined with pooling of the appropriate gradient fractions. These two subtypes differ in their dependence on the cortical actin cytoskeleton.