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. (see product sheet for more info)

Product Attachments

View FileUbiQ-073 Productsheet B15112014-001
DUB activity profilingcompound library screening

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name ISG15-FP
product code UbiQ-073
Applications DUB activity profiling, compound library screening
target Ubl proteases
source mouse sequence, semi-synthetic
shipping lyophilized powder
purity >95%
molecular weight 18.25 kDa
storage upon arrival, powder at −20°, solution at −80°C. Please protect from light and avoid multiple freeze/thaw cycles.
sample preparation For detailed sample preparation, please see product sheet.
regulatory statement For laboratory research use only, not for use in humans.
  • A. Tirat et al.

    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).

  • Huang et al.

    Huang, X. & Aulabaugh A. Application of fluorescence polarization in HTS assays. Methods in Molecular Biology 565, 127-143 (2009).

  • Levine et al.

    Levine, L.M., et al. Measurement of specific protease activity utilizing fluorescence polarization. Anal. Biochem. 247, 83-88 (1997).

  • Geurink and El Oualid et al.

    Geurink, P.P., et al. A general chemical ligation approach towards isopeptide-linked ubiquitin and ubiquitin-like assay reagents. ChemBiochem 13, 293-297 (2012).

  • Basters et al.

    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).

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