Isolation of GLUT4 storage vesicles.

The immunoisolation of GLUT4-containing vesicles from adipocytes is described in this unit. The methods involve homogenization of cells followed by differential centrifugation to provide the intracellular membranes that contain GLUT4. Subsequently, an immobilized monoclonal antibody is used for the isolation of vesicles of very high purity. The various protocols are applicable to cultured and primary adipocytes as well as skeletal muscle, the major insulin target cells expressing GLUT4.