Jr. Petrulis et Gh. Perdew, Monitoring nuclear import with GFP-variant fusion proteins in digitonin-permeabilized cells, BIOTECHNIQU, 31(4), 2001, pp. 772-775
A convenient assay for monitoring nuclear localization signal-mediated nucl
ear import of green fluorescent protein (GFP)-variant fusion proteins has b
een developed. This modified assay relies upon indirect immunofluorescence
microscopy for visualization of transported substrates. The use of GFP-vari
ant fusion proteins allows for the rapid assessment of optimal digitonin co
ncentration and permits nuclear import to be monitored with minimal sample
preparation in real time.