1974/09/01 by H. Heitzmann, Frederic M. Richards · 3 citations
Biochemistry, Genetics and Molecular Biology · Engineering · Chemistry · #Biotin and Related Studies #Lipid Membrane Structure and Behavior #Electrochemical sensors and biosensors #Avidin #Biotin #Biochemistry #Staining #Ferritin #Galactose oxidase #Chemistry #Membrane #Electron microscope #Galactose #Biology
paper · doi:10.1073/pnas.71.9.3537
openalex publication_date 1974/09/01 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/15
To expand the electron microscopist's options in localization and visualization, a new and general staining technique has been tested. The avidin-biotin complex serves as a coupling between the electron-dense marker, ferritin, and points of interest in biological samples. When specific cellular components are tagged with biotin, those components may be visualized with ferritin-linked avidin. Because of the remarkably strong affinity of avidin and biotin (characterized by an association constant of 10(15) M(-1)), the staining is rapid and stable. The preparation of ferritin-avidin conjugate is described, and examples are presented of the application of this complex to biotin-tagged membranes. The ghosts of Acholeplasma laidlawii have been treated with biotinyl-N-hydroxysuccinimide ester to label protein amino groups. Erythrocyte membrane oligosaccharides have been oxidized by periodate or by galactose oxidase, and the resulting aldehydes labeled with biotin hydrazide. The avidin-biotin complex in electron microscopy seems especially appropriate for seqential staining procedures, as well as for visualization of reaction sites of biotin-labeled, low-molecular-weight reagents.