title: Preparation of detergent-resistant membranes (DRMs) from cultured mammalian cells.
abstract: Detergent-resistant membranes (DRMs) isolated from cells are enriched in proteins and lipids with a high affinity for lipid rafts, or membrane microdomains in the liquid-ordered phase. Enrichment in DRMs provides a good indication that a protein is "raftophilic," and may be present in rafts in cell membranes before extraction. Here, I describe preparation of Triton X-100-insoluble DRMs from cultured mammalian cells on sucrose gradients. Methods for analyzing the distribution of particular proteins across the gradient, and for recovering DRMs for further use are presented.
authors.affiliation: Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, NY, 11794-5215, USA, Deborah.brown@stonybrook.edu.
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Brown
mesh_terms: D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000187:drug effects*; D002478:Cells, Cultured; D003902:Detergents / Q000494:pharmacology*; D008322:Mammals; D008967:Molecular Biology / Q000379:methods*; D013395:Sucrose; D014461:Ultracentrifugation
chemical_list: D003902:Detergents; D013395:Sucrose