Peer-Reviewed Protocol | Open Access
Separation of Membrane Vesicles and Cytosol from Cultured Cells and Bacteria in a Preformed Discontinuous Gradient
There are many situations when it is necessary to separate rapidly and efficiently a cytosolic and a membrane vesicle fraction from either cultured cells or from bacteria. Flotation of the vesicles through a low-density barrier from a dense sample zone using the low viscosity medium iodixanol allows complete separation of these compartments. As the sample is exposed to the gmax the tendency of the proteins to sediment overcomes any diffusion in the opposite direction.