Use of the biotin-avidin system for labelling, isolation and characterization of neural cell-surface proteins.
We describe a method for the selective labelling, isolation and electrophoretic analysis of cell-surface molecules and extracellular matrix components. Intact tissues are reacted with activated esters of biotin and the labelled surface molecules identified on Western blots with horseradish-peroxidase-coupled or 35S-labelled streptavidin. Alternatively, the biotinylated proteins can be purified from tissue homogenates by affinity chromatography on an avidin-agarose column. Evidence is presented to show that this method is indeed specific for membrane and matrix components. Its practical application to embryonic neural tissues is demonstrated.
von Boxberg
Y
Y
Max-Planck-Institut für Entwicklungsbiologie, Abteilung Biochemie, Tübingen, Federal Republic of Germany.
Wütz
R
R
Schwarz
U
U
D000818:Animals; D001360:Avidin; D001369:Axons / Q000032:analysis; D001710:Biotin; D002642:Chick Embryo; D002645:Chickens; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006651:Histocytochemistry; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000032:analysis* / Q000648:ultrastructure; D008854:Microscopy, Electron; D012160:Retina / Q000032:analysis* / Q000196:embryology / Q000648:ultrastructure; D013194:Staining and Labeling
D008565:Membrane Proteins; D001360:Avidin; D001710:Biotin