Monitoring Nuclear Import with GFP-Variant Fusion Proteins in Digitonin-Permeabilized Cells

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-vari...

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Veröffentlicht in:	BioTechniques 2001-10, Vol.31 (4), p.772-775
Hauptverfasser:	Petrulis, John R, Perdew, Gary H
Format:	Artikel
Sprache:	eng
Schlagworte:	Active Transport, Cell Nucleus Analytical, structural and metabolic biochemistry Animals Biological and medical sciences Biotechnology Cell Membrane Permeability COS Cells Digitonin Fundamental and applied biological sciences. Psychology Gene transfer General aspects, investigation methods Genetic engineering Genetic technics Genetic Variation Green Fluorescent Proteins Luminescent Proteins - genetics Luminescent Proteins - metabolism Methods. Procedures. Technologies Microscopy, Fluorescence Proteins Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Synthetic digonucleotides and genes. Sequencing
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Beschreibung
Zusammenfassung:	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.
ISSN:	0736-6205 1940-9818
DOI:	10.2144/01314st02