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Localization and quantification of epidermal growth factor receptors on single cells by confocal laser scanning microscopy
MJ Good, WJ Hage, CL Mummery, SW De Laat and J Boonstra
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, University of Utrecht.
We have established a method for quantifying binding of fluorescence- labeled growth factors to their receptors on single cells in situ with the confocal laser scanning microscope (CLSM). Biotinylated epidermal growth factor (EGF) coupled to phycoerythrin-labeled anti-biotin was used to compare the levels of fluorescence on three different cell types for which the number of EGF factors was known from Scatchard analysis of [125I]-EGF binding. The results showed that as few as 10,000 receptors/cell were detectable above back-ground. This method will provide a rapid and quantifiable alternative to autoradiography for ligand binding to single cells in situ.
Volume 40,
Issue 9,
pp. 1353-1361,
09/01/1992
Copyright © 1992 by The Histochemical Society
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