Purification of brush border membrane by thiocyanate treatment.
Rat small intestinal brush border membranes are purified from brush borders by homogenization in relatively high concentrations of thiocyanate salts (0.56 M LiSCN, 0.41 M NaSCN, or 0.52 M KSCN), removal of this salt, and differential centrifugation to separate cytoskeletal material from membranes. The marker enzyme, sucrase, is enriched 98-fold in the final membranes over the starting homogenate of intestinal scrapings at a yield of about 20%. The isolated membranes are capable of secondary active sodium-dependent glucose transport as demonstrated by sodium gradient-supported overshooting glucose uptake.
D000818:Animals; D001692:Biological Transport; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000648:ultrastructure; D005947:Glucose / Q000378:metabolism; D007421:Intestine, Small / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D008871:Microvilli / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D011919:Rats, Inbred Strains; D013861:Thiocyanates
D013861:Thiocyanates; D005947:Glucose