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Studying Purinoceptor Cell-Surface Expression by Protein Biotinylation

  • Mark T. YoungEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2041)

Abstract

Covalent labeling of protein with biotin (biotinylation) is a versatile technique which enables the capture and analysis of labeled protein with streptavidin-coated beads. It is particularly useful for detecting and quantifying the cell-surface expression of membrane proteins and widely used to analyze protein trafficking and the effect of mutations (particularly those which render the protein nonfunctional) on cell-surface expression. Here I describe the procedure for biotinylation and capture of cell-surface rat P2X2 receptors expressed in mammalian cells, and outline the steps in data analysis required to measure the proportion of cell-surface expressed protein of single point mutants relative to a wild-type control.

Key words

Cell-surface Protein expression Biotinylation P2X Western blot Crosslinking Membrane protein 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.School of BiosciencesCardiff UniversityCardiffUK

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