Localization and quantification of epidermal growth factor receptors on single cells by confocal laser scanning microscopy.
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.
Good
M J
MJ
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, University of Utrecht.
Hage
W J
WJ
Mummery
C L
CL
De Laat
S W
SW
Boonstra
J
J
D001710:Biotin / Q000378:metabolism; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism* / Q000648:ultrastructure; D005453:Fluorescence; D007834:Lasers; D008854:Microscopy, Electron / Q000379:methods
D004815:Epidermal Growth Factor; D001710:Biotin; D066246:ErbB Receptors