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High-throughput analysis of the dynamics of recycling cell surface proteins


Recycling via the plasma membrane is a key feature that is shared by many membrane proteins. Using a combination of indirect immunofluorescence labeling and fluorescence detection using a fluorescence multiwell plate reader, we exploited the possibilities of quantitatively measuring the trafficking kinetics of transmembrane proteins. Parameters that can be studied include dynamic appearance/presence at the cell surface, recycling via the cell surface, and internalization. For the insulin-responsive glucose transporter GLUT4 (glucose transporter number 4), details are presented on how to quantitatively measure insulin-induced GLUT4 translocation toward the plasma membrane (transition state) and to analyze cell surface recycling of GLUT4 in basal and insulin-stimulated cells (steady state).

Type Journal
ISBN 1064-3745 (Print)
Authors Govers, R.;James, D. E.;Coster, A. C. :
Publisher Name Methods in Molecular Biology
Published Date 2008-01-01
Published Volume 440
Published Pages 129-46
Status Published in-print
URL link to publisher's version
OpenAccess link to author's accepted manuscript version