Fractionation of Golgi, endoplasmic reticulum, and plasma membrane from cultured cells in a preformed continuous iodixanol gradient.
A continuous iodixanol gradient within the range 0-30% (w/v) iodixanol can resolve the major membrane compartments of the endoplasmic reticulum, Golgi membranes, and plasma membrane from a postnuclear supernatant prepared from a cultured cell homogenate. The precise density range of the gradient and the centrifugation conditions (100,000-200,000 g for 2-16 h) vary with the type of cell and the requirements of the separation. The strategy is widely used to study the processing of proteins within cells.
Graham
John M
JM
School of Biomolecular Sciences, Liverpool John Moores University. john@jgrescon.fsbusiness.co.uk
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D019556:COS Cells / Q000737:chemistry; D002458:Cell Fractionation / Q000379:methods; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002500:Centrifugation, Isopycnic / Q000379:methods*; D002522:Chlorocebus aethiops; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000378:metabolism*; D006056:Golgi Apparatus / Q000737:chemistry / Q000378:metabolism*; D014283:Triiodobenzoic Acids / Q000737:chemistry*
D015415:Biomarkers; D014283:Triiodobenzoic Acids; C044834:iodixanol