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-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.
Petrulis
J R
JR
The Pennsylvania State University, University Park 16802, USA.
Perdew
G H
GH
D021581:Active Transport, Cell Nucleus; D000818:Animals; D001709:Biotechnology; D019556:COS Cells; D002463:Cell Membrane Permeability; D004072:Digitonin; D014644:Genetic Variation; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D004072:Digitonin