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

John R. Petrulis | Zendy; Gary H. Perdew | Zendy

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

Author(s) -

John R. Petrulis,

Gary H. Perdew

Publication year - 2001

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/01314st02

Subject(s) - green fluorescent protein , digitonin , fusion protein , nuclear transport , nuclear localization sequence , immunofluorescence , recombinant dna , fluorescence , fluorescence microscope , microbiology and biotechnology , biology , chemistry , biochemistry , cell nucleus , enzyme , gene , antibody , genetics , cytoplasm , physics , quantum mechanics

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.

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