Demonstration of a Na+/H+ exchanger NHE1 in fresh bovine corneal endothelial cell basolateral plasma membrane

Apical and basolateral plasma membranes of fresh bovine corneal endothelial cells were isolated using positively charged polyacrylamide beads. Marker enzyme assays demonstrated that the isolated apical and basolateral plasma membrane domains could be isolated and separated with relative purity. West ern blotting with a polyclonal anti-NHE1 antibody detected a protein of 70 kDa in the basolateral plasma membrane isolate. NHE1 immunoreactivity was n ot detected in the apical membrane sample. This suggests that the Na+/H+ ex changer, NHE1, is strictly localised to the basolateral membrane of fresh b ovine corneal endothelial cells. (C) 1999 Published by Elsevier Science B.V . All rights reserved.