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Mol Biosyst. 2013 Mar;9(3):339-42. doi: 10.1039/c3mb25552b. Epub 2013 Jan 22.

Fluorogenic label to quantify the cytosolic delivery of macromolecules.

Author information

1
Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53706, USA.

Abstract

The delivery of a macromolecule to the cytosol of human cells is assessed by using a pendant di-O-glycosylated derivative of fluorescein. Its fluorescence is unmasked by Escherichia coliβ-galactosidase installed in the cytosol. Background is diminished by using RNAi to suppress the expression of GLB1, which encodes a lysosomal β-galactosidase. This strategy was used to quantify the cytosolic entry of a highly cationic protein, ribonuclease A.

PMID:
23340874
PMCID:
PMC3588161
DOI:
10.1039/c3mb25552b
[Indexed for MEDLINE]
Free PMC Article

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