Fluorogenic label to quantify the cytosolic delivery of macromolecules

被引：20
|
作者：
Chao, Tzu-Yuan [1 ]
Raines, Ronald T. [1 ,2 ]
机构：
[1] Univ Wisconsin, Dept Biochem, Madison, WI 53706 USA
[2] Univ Wisconsin, Dept Chem, Madison, WI 53706 USA
来源：
MOLECULAR BIOSYSTEMS | 2013年 / 9卷 / 03期
关键词：
BETA-GALACTOSIDASE; RIBONUCLEASE INHIBITOR; FLOW-CYTOMETRY; FLUORESCEIN; VARIANTS; PROTEINS; DESIGN;
D O I：
10.1039/c3mb25552b
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
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 coli beta-galactosidase installed in the cytosol. Background is diminished by using RNAi to suppress the expression of GLB1, which encodes a lysosomal beta-galactosidase. This strategy was used to quantify the cytosolic entry of a highly cationic protein, ribonuclease A.
引用
收藏
页码：339 / 342
页数：4
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