ARTICLE

THE LOCALIZATION OF SPECTRIN ON THE INNER SURFACE OF HUMAN RED BLOOD CELL MEMBRANES BY FERRITIN-CONJUGATED ANTIBODIES

Garth L. Nicolson ¹, V. T. Marchesi ¹, and S. J. Singer ¹

¹ From the Department of Biology, University of California at San Diego, La Jolla, California 92037, and the National Institute of Arthritis and Metabolic Diseases, National Institutes of Health, Bethesda, Maryland 20014.

Dr. Nicolson's present address is the Armand Hammer Cancer Center of The Salk Institute for Biological Studies, La Jolla, California 92037

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 ß-mercaptoethanol prevented labeling by specific ferritin-conjugated antibodies.

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This article has been cited by other articles:

• Bennett, V., Davis, J. (1982). Immunoreactive Forms of Human Erythrocyte Ankyrin Are Localized in Mitotic Structures in Cultured Cells and Are Associated with Microtubules in Brain. Cold Spring Harb Symp Quant Biol 46: 647-657 [Abstract]  
• Nicolson, G. L., Yanagimachi, R. (1974). Mobility and the Restriction of Mobility of Plasma Membrane Lectin-Binding Components. Science 184: 1294-1296 [Abstract]  
• Bretscher, M. S. (1973). Membrane Structure: Some General Principles. Science 181: 622-629 [Abstract]  
• Nicolson, G. L., Yanagimachi, R. (1972). Terminal Saccharides on Sperm Plasma Membranes: Identification by Specific Agglutinins. Science 177: 276-279 [Abstract]  

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