ISG15-FP

a fluorescence polarization assay reagent for deISGylating enzymes, based on a TAMRA modified Lys-Gly sequence, linked to mouse ISG15

productsheet

code UbiQ-073 Category
product UbiQ-073 Category

Additional information

Weight 0.05 kg
bulk size

, ,

aliquot size

Applications

,

target

,

source

shipping

purity

>95%

molecular weight

storage

sample preparation

For detailed sample preparation, please see product sheet.

regulatory statement

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300.00

Description

UbiQ-073 is a fluorescence polarization assay reagent for deISGylating enzymes which is based on a 5-carboxytetramethylrhodamine (TAMRA, exc 550 nm, emi 590 nm) modified Lys-Gly sequence that is linked to (mouse) ISG15 (Interferon stimulated gene 15) via a native isopeptide bond with the lysine side-chain. ISG15 is a ubiquitin-like protein whose expression is increased following stimulation with type 1 Interferons.

A typical substrate concentration is 100 nM. Effective concentrations of deISGylating enzymes depends on specific assay conditions and method of detection.

UbiQ-073 is effectively processed by the known deISGylating enzyme USP18.

Additional information

Weight 0.05 kg
bulk size

, ,

aliquot size

Applications

,

target

,

source

shipping

purity

>95%

molecular weight

storage

sample preparation

For detailed sample preparation, please see product sheet.

regulatory statement

Tirat, A., et al. Synthesis and characterization of fluorescent ubiquitin derivatives as highly sensitive substrates for the deubiquitinating enzymes UCH-L3 and USP-2. Anal. Biochem. 343, 244-255 (2005).
http://www.ncbi.nlm.nih.gov/pubmed/15963938

Huang, X. & Aulabaugh A. Application of fluorescence polarization in HTS assays. Methods in Molecular Biology 565, 127-143 (2009).
http://www.ncbi.nlm.nih.gov/pubmed/19551360

Levine, L.M., et al. Measurement of specific protease activity utilizing fluorescence polarization. Anal. Biochem. 247, 83-88 (1997).
http://www.ncbi.nlm.nih.gov/pubmed/9126375

Geurink, P.P., et al. A general chemical ligation approach towards isopeptide-linked ubiquitin and ubiquitin-like assay reagents. ChemBiochem 13, 293-297 (2012).
http://www.ncbi.nlm.nih.gov/pubmed/22213387

Basters, A., et al. Molecular characterization of ubiquitin-specific protease 18 reveals substrate specificity for interferon-stimulated gene 15. Febs. J. 281, 1918-1928 (2014).
http://www.ncbi.nlm.nih.gov/pubmed/24533902