A detergent-free method for purifying caveolae membrane from tissue culture cells.
Current methods for purifying caveolae from tissue culture cells take advantage of the Triton X-100 insolubility of this membrane domain. To circumvent the use of detergents, we have developed a method that depends upon the unique buoyant density of caveolae membrane. The caveolae fractions that we obtain are highly enriched in caveolin. As a consequence we are able to identify caveolae-associated proteins that had previously gone undetected. Moreover, resident caveolae proteins that are soluble in Triton X-100 are retained during the isolation.
D015415:Biomarkers; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000648:ultrastructure*; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004798:Enzymes / Q000032:analysis; D005347:Fibroblasts / Q000166:cytology / Q000648:ultrastructure; D006801:Humans; D015151:Immunoblotting; D008854:Microscopy, Electron; D015388:Organelles / Q000648:ultrastructure*; D011506:Proteins / Q000032:analysis; D012867:Skin / Q000166:cytology / Q000648:ultrastructure*; D014461:Ultracentrifugation
D015415:Biomarkers; D004798:Enzymes; D011506:Proteins