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.
CITATION STYLE
Young, M. T. (2020). Studying purinoceptor cell-surface expression by protein biotinylation. In Methods in Molecular Biology (Vol. 2041, pp. 137–146). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9717-6_9
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