Abstract
A convenient assay for monitoring nuclear localization signal-mediated nuclear import of green fluorescent protein (GFP)-variant fusion proteins has been developed. This modified assay relies upon indirect immunofluorescence microscopy for visualization of transported substrates. The use of GFP-variant fusion proteins allows for the rapid assessment of optimal digitonin concentration and permits nuclear import to be monitored with minimal sample preparation in real time.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Active Transport, Cell Nucleus*
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Animals
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Biotechnology
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COS Cells
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Cell Membrane Permeability
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Digitonin
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Genetic Variation
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Green Fluorescent Proteins
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism*
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Microscopy, Fluorescence
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
Substances
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Luminescent Proteins
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Recombinant Fusion Proteins
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Green Fluorescent Proteins
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Digitonin