PMID: 3137189Jan 1, 1988

Labelling of colloidal gold with IgE. A quantitative study using monoclonal IgE anti-beta-lactoglobulin and evaluation of the biological activity of the gold complex with RBL-1 cells

Histochemistry
M F ClercM Horisberger

Abstract

Immunocytochemical markers prepared by labelling colloidal gold with antibodies are gaining wide acceptance both in transmission and scanning electron microscopy. However, detailed information on the process and extent of adsorption of IgG and IgE in particular are still lacking. The adsorption isotherm of mouse monoclonal 125I-IgE antibovine milk beta-lactoglobulin was studied quantitatively with colloidal gold buffered at pH 6.1-8.8 (28 nm in particle diameter). At low coverage of the particles (less that or equal to 5 molecules per particle), the isotherm was independent of pH. In the presence of a large excess of IgE, the highest coverage was obtained at pH 6.1 near the pI of IgE (5.2-5.8). The binding constants were higher at low coverage (side-on adsorption) than at high coverage where desorption was observed. IgE-Au markers were unreactive towards the immobilized antigen and did not bind to receptors for IgE of rat basophilic leukemia cells (RBL-1). The reactivity of immobilized anti-IgE antibodies with IgE-Au markers increased as a function of particle coverage. Mapping of RBL-1 cell membrane IgE receptors was achieved by incubating successively IgE-sensitized RBL-1 cells with anti-IgE antibodies and a protein A-gold ma...Continue Reading

References

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Citations

Jan 1, 1995·Progress in Histochemistry and Cytochemistry·M Bendayan
Jul 1, 1996·The Journal of Laboratory and Clinical Medicine·S R Simmons, R M Albrecht
Dec 31, 1997·Arteriosclerosis, Thrombosis, and Vascular Biology·S R SimmonsR M Albrecht
Aug 19, 2007·Molecular Biology of the Cell·Sophie G MartinFred Chang

Related Concepts

Monoclonal Antibodies
Hydrocolloids
Gold
Immunoglobulin epsilon-Chains
Immunoglobulin Heavy Chain Subgroup VH-III
Lactoglobulins
Receptors, Immunologic

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