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Laterobasal membranes from intestinal epithelial cells: Isolation free of intracellular membrane contaminants |
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| Authors: | Toan D Nguyen Jean-Pierre Broyart Khoanh T Ngu Annabella Illescas Austin K Mircheff Gary M Gray |
| Institution: | (1) Division of Gastroenterology, Department of Medicine, Stanford University, 94305 Stanford, California;(2) Department of Physiology, University of Southern California, 90033 Los Angeles, California;(3) Present address: V. A. Hospital, 508 Fulton Street, Building 2, Floor 2, 27705 Durham, NC;(4) Present address: INSERM U-120, 44 Chemin de Ronde, 7810 Le Vesinet, France |
| Abstract: | Summary A simplified method for isolating highly purified laterobasal membranes (LBM) from enterocytes is based on treatment of membranes with 8mm CaCl2 concentration in order to aggregate intracellular membrane contaminants. The resultant LBM showed an average 15-fold enrichment and constituted 8% of the original K-stimulated phosphatase in the initial crude homogenate. It showed typical LBM migration on counter-current distribution (CCD) and was essentially free of contamination with endoplasmic reticulum and Golgi membranes. This method is highly efficient and yields sufficient purified LBM to allow comprehensive analysis of enterocyte membrane events. |
| Keywords: | small intestine enterocyte membrane fractionation laterobasal membrane endoplasmic reticulum Golgi |
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