title: Biotin-ligand complexes with streptavidin quantum dots for in vivo cell labeling of membrane receptors.
abstract: The unique fluorescence properties of quantum dots (QDs), particularly their large extinction coefficients and photostability, make them ideal probes for tracking proteins in live cells using real-time visualization. We have shown that QDs conjugated to epidermal growth factor act as functional ligands for their receptor, erbB1. Here, we describe protocols for (1) conjugation of streptavidin-QDs to biotinylated ligand, (2) formation of ligand-QD-receptor complexes, and (3) quantification of binding and internalization of receptor complex using both high-resolution fluorescence microscopy and flow cytometry.
authors.affiliation: Department of Pathology, Univesity of New Mexico, Albuquerque, NM, USA.
authors.forename: Diane S
authors.initials: DS
authors.lastname: Lidke
authors.forename: Peter
authors.initials: P
authors.lastname: Nagy
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Jovin
authors.forename: Donna J
authors.initials: DJ
authors.lastname: Arndt-Jovin
mesh_terms: D000818:Animals; D001710:Biotin / Q000737:chemistry* / Q000378:metabolism; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000378:metabolism; D066246:ErbB Receptors / Q000737:chemistry / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D008024:Ligands; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D045663:Quantum Dots; D019809:Streptavidin / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008024:Ligands; D004815:Epidermal Growth Factor; D001710:Biotin; D019809:Streptavidin; D066246:ErbB Receptors