Selective immunocytochemical detection of fluorescent analogs with antibodies specific for the fluorophore.

K. Luby-Phelps, P. A. Amato, D. L. Taylor

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Fluorescent analogs of cellular components are finding increasing use in the field of cell biology. The power of this technique can be augmented by the use of antibodies specific for the fluorophore to visualize selectively the fluorescent analog at the electron microscope level. Rabbit antibodies specific for fluorescein were elicited and purified according to published methods (Lopatin and Voss [1971]: Biochemistry 10:208). Immune sera and IgG formed precipitin lines with fluorescein-labeled proteins in Ouchterlony immunodiffusion assays, and significantly significantly quenched the fluorescence of fluorescein-labeled proteins. Immune IgG and Fab fragments decorated fluorescein-labeled actin, but not unlabeled actin, in negative-stained preparations. Anti-fluorescein IgG was used for immunofluorescent localization of fluorescein-labeled actin following microinjection of the fluorescent analog into living cells. This approach was extended to the immunoelectron microscopic localization of the injected analog at the subcellular level by the use of an electron-dense marker coupled to goat anti-rabbit IgG. Many other fluorescent probes also can be used as haptens for production of antibodies. Therefore, a general method for localizing fluorescently labeled molecules at the electron microscopic level is now available. Several other applications of anti-fluorescein antibody in studies involving fluorescent analogs are also suggested.

Original languageEnglish (US)
Pages (from-to)137-149
Number of pages13
JournalCell Motility
Volume4
Issue number2
StatePublished - 1984

Fingerprint

Fluorescein
Antibodies
Actins
Electrons
Immunoglobulin G
Rabbits
Precipitins
Immunoglobulin Fab Fragments
Haptens
Immunodiffusion
Microinjections
Fluorescent Dyes
Goats
Biochemistry
Antibody Formation
Cell Biology
Immune Sera
Anti-Idiotypic Antibodies
Proteins
Fluorescence

ASJC Scopus subject areas

  • Cell Biology

Cite this

Selective immunocytochemical detection of fluorescent analogs with antibodies specific for the fluorophore. / Luby-Phelps, K.; Amato, P. A.; Taylor, D. L.

In: Cell Motility, Vol. 4, No. 2, 1984, p. 137-149.

Research output: Contribution to journalArticle

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