The localization of spectrin on the inner surface of human red blood cell membranes by ferritin-conjugated antibodies.
Spectrin, a major protein constituent of mammalian red blood cell membrane preparations, has been localized on the inner surface of human red blood cell membranes by techniques that utilized specific ferritin-conjugated antibodies and fixation of membranes shortly after hemolysis so as to allow penetration of the ferritin-antibody labels. The labeling of spectrin was shown to be specific by the following criteria. (a) Nonhomologous ferritin-conjugated antibodies did not specifically bind to either membrane surface. (b) Blocking the membrane-bound spectrin with excess unconjugated antispectrin antibodies prevented ferritin-antibody labeling. (c) Removal of spectrin by treating the membrane preparation with a low ionic strength buffer containing ethylenediaminetetraacetate and beta-mercaptoethanol prevented labeling by specific ferritin-conjugated antibodies.
D000818:Animals; D000906:Antibodies; D000917:Antibody Formation; D001798:Blood Proteins; D002021:Buffers; D002462:Cell Membrane / Q000276:immunology; D002848:Chromatography, DEAE-Cellulose; D004492:Edetic Acid; D004912:Erythrocytes / Q000276:immunology; D005293:Ferritins; D006461:Hemolysis; D006651:Histocytochemistry; D006736:Horses; D006801:Humans; D007106:Immune Sera; D007552:Isotonic Solutions; D008623:Mercaptoethanol; D008854:Microscopy, Electron; D009975:Orthomyxoviridae; D011485:Protein Binding; D011817:Rabbits
D000906:Antibodies; D001798:Blood Proteins; D002021:Buffers; D007106:Immune Sera; D007552:Isotonic Solutions; D008623:Mercaptoethanol; D005293:Ferritins; D004492:Edetic Acid