Labelling of colloidal gold with protein
- 1 January 1984
- journal article
- research article
- Published by Springer Nature in Histochemistry and Cell Biology
- Vol. 80 (1), 13-18
- https://doi.org/10.1007/bf00492765
Abstract
Markers prepared by labelling colloidal gold with macromolecules such as lectins, antibodies and protein A are gaining wise acceptance both in transmission and scanning electron microscopy. However detailed information on the process and extent of adsorption of macromolecules onto gold particles are still lacking. The adsorption isotherm of protein onto gold particles was studied quantitatively using goat β-lactoglobulin (βL) tritiated in vivo. When this protein was modified chemically by iodination with 125I, the adsorption isotherm was not significantly different (Langmuir type for monolayer). In the presence of saturating amount of βL, a maximum of 13–14 molecules was adsorbed per particle of 12 nm in diameter for a theoretical maximum of 20 (compact monolayer). Ellipsometric measurements on nickel-coated slides indicated that βL was adsorbed onto metallic surfaces as a compact monolayer. The molecules were irreversibly adsorbed on gold particles, kept to a large extent their capacity to bind anti-βL antibodies and could not be displaced by polyethylene glycol, a stabilizer commonly used in the preparation of gold markers. Only markers labelled with more than 5 βL molecules per particle could be completely bound by immobilized anti-βL antibodies. Preliminary data indicated that the energetics of adsorption of βL onto colloidal gold was in agreement with that expected from the mutual interaction of surface and adsorbate.Keywords
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