Laterobasal membranes from intestinal epithelial cells: isolation free of intracellular membrane contaminants.
A simplified method for isolating highly purified laterobasal membranes (LBM) from enterocytes is based on treatment of membranes with 8 mM 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.
Nguyen
T D
TD
Department of Medicine, Stanford University, California 94305.
Broyart
J P
JP
Ngu
K T
KT
Illescas
A
A
Mircheff
A K
AK
Gray
G M
GM
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000648:ultrastructure*; D002499:Centrifugation, Density Gradient; D003377:Countercurrent Distribution; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D004848:Epithelium / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000648:ultrastructure; D007082:Ileum / Q000648:ultrastructure*; D007413:Intestinal Mucosa / Q000648:ultrastructure*; D007425:Intracellular Membranes / Q000648:ultrastructure; D007583:Jejunum / Q000648:ultrastructure*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains