A new method for the isolation of renal basement membranes.
A method is described for the isolation of basement membranes from rabbit renal cortex in which the detergent N-lauroyl sarcosine is used as the disruptive agent. The isolated membranes have been compared with membranes prepared using ultrasonication and they were comparable both in terms of purity and gross chemical composition. Glomerular and tubular basement membranes were isolated by first separating glomeruli from tubules by density gradient centrifugation followed by detergent treatment of the separated tissues. The detergent method has the advantage that the basement membranes retained their native structure to a large degree, whereas sonicated membranes were severely fragmented. Collagen fibres were a significant contaminant in both preparations and were revealed more clearly by negative staining than by examination of thin sections. Studies with the detergent-treated membrane revealed that a few proteins, which seemed to be membrane components, were extracted with 1 M NaCl and that these proteins were lost from the basement membranes during sonication used in the conventional isolation procedure.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001485:Basement Membrane / Q000648:ultrastructure*; D003902:Detergents; D005260:Female; D007672:Kidney Cortex / Q000648:ultrastructure*; D007850:Lauric Acids; D008297:Male; D008565:Membrane Proteins / Q000032:analysis; D008722:Methods; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D008970:Molecular Weight; D011817:Rabbits; D012521:Sarcosine; D014465:Ultrasonics
D000596:Amino Acids; D003902:Detergents; D007850:Lauric Acids; D008565:Membrane Proteins; D012521:Sarcosine