A Covalent Fluorescent–Gold Immunoprobe: Simultaneous Detection of a Pre-mRNA Splicing Factor by Light and Electron Microscopy
Year: 1997
Type: article
Abstract: Immunoprobes that combine a fluorescent label with a 1.4-nm gold cluster compound have been prepared by covalent conjugation with polyclonal antibody Fab' fragments. These new immunoconjugates allow the collection of two complementary sets of data, from fluorescence and electron microscopy, from a single labeling experiment. We find that incorporation of one or more fluorescein moieties into the coordinated ligands of the 1.4-nm Nanogold gold cluster label yields a stable, dual-function immunolabel in which fluorescence quenching is negligible. In a second synthetic strategy, Nanogold and fluorescein were separately covalently conjugated to Fab' fragments to yield a probe with very similar properties. With the use of Fab' fragments, the entire probe is smaller than a whole IgG molecule, and it exhibited excellent penetration properties. It was used to localize the pre-mRNA splicing factor SC35 in the HeLa cell nucleus by both fluorescence and electron microscopy.
Authors: Richard D. Powell, Carol M. R. Halsey, David L. Spector, Shelley L. Kaurin, Jennifer McCann, James F. Hainfeld
Institution: Cold Spring Harbor Laboratory
Language: English
Subfield: Materials Chemistry
Field: Materials Science
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $4,400
Award: G4217455733
OpenAlex ID: W2018428295