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I9906

Sigma-Aldrich

Import Ligand, Fluorescent

buffered aqueous solution

Synonym(s):
Sulforhodamine B Labeled Albumin-NLS Peptide
MDL number:

sterility

sterile; 0.2 μm filtered

form

buffered aqueous solution

composition

Dye content, 2-4 mol/mol protein (rhodamine B/bovine serum albumin)
NLS peptide, 8-20 mol/mol protein (NLS/bovine serum albumin)

color

red to purple

fluorescence

λex 560 nm; λem 580 nm

shipped in

dry ice

storage temp.

−20°C

Biochem/physiol Actions

Common NLS (nuclear localization signal) for semi-synthetic import ligands is the SV-40 large T-antigen NLS, amino acid sequence CGGGPKKKRKVED covalently linked to rhodamine-labeled albumin. Enables observation of the nuclear import process in permeabilized cells by fluorescence microscopy.
Proteins due to enter the cell nucleus are ‘labeled′ by nuclear localization signals (NLS). The canonical NLS consists of either a single or bipartite stretch of primarily basic amino acids. The NLS is recognized in the cyto­plasm by its receptor importin α. Importin α forms a complex with another protein, importin β. This inter­action then targets the import complex to the nuclear pore complex (NPC). Passage of the import substrate from the NPC to the nuclear interior requires two ad­di­tion­al factors: p10 and the small GTPase Ran. In the nucleus, the binding of Ran-GTP to importin β releases the transported protein from the complex.
A commonly used NLS (nuclear localization signal) for semi-synthetic import ligands is the SV-40 large T-antigen NLS, a 13 amino acid peptide of the sequence CGGGPKKKRKVED, which is covalently linked to rhodamine-labeled albumin. This enables observation of the nuclear import process in permeabilized cells by fluorescence microscopy using a filter suitable for TRITC/Cy3.

Analysis Note

Optimal import into nuclei of digitonin-permeabilized HeLa cells is observed at a concentration of 120 μg/ml.

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