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Single-Cell Quantification of Protein Degradation Rates by Time-Lapse Fluorescence Microscopy in Adherent Cell Culture
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February 04, 2018
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This protocol describes a method to determine protein half-lives in single living adherent cells, using pulse labeling and fluorescence time-lapse imaging of SNAP-tag fusion proteins.
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Protein DegradationSingle-cellTime-lapse Fluorescence MicroscopyAdherent Cell CultureSNAP-tagProtein Half-lifeFluorescence ImagingProtein TurnoverHeterogeneityEndogenous Protein TaggingOverexpressionDNA Repair EnzymeBenzylguanineFluorescent DyeExponential DecayEmbryonic Stem CellsMonolayerCell-cell InteractionsN-cadherin
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Alber, A. B., Suter, D. M. Single-Cell Quantification of Protein Degradation Rates by Time-Lapse Fluorescence Microscopy in Adherent Cell Culture. J. Vis. Exp. (132), e56604, doi:10.3791/56604 (2018).
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