Nedd8-FP (UbiQ-019) is a fluorescence polarization assay reagent which is based on a 5-carboxytetramethylrhodamine (TAMRA) modified Lys-Gly sequence that is linked to Nedd8 via a native isopeptide bond with the lysine side-chain. Nedd8 protease concentrations can range from 0.01-10 nM but depend on specific assay conditions and method of detection. Typical substrate concentrations range from 10−100 nM.

Product Attachments

View FileUbiQ-019 Productsheet B01052014-001
DUB activity profilingcompound library screening

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name NEDD8-FP
product code UbiQ-019
Applications DUB activity profiling, compound library screening
target Ubl proteases
source human sequence, semi-synthetic
shipping lyophilized powder
purity >95%
molecular weight 9.16 kDa
storage upon arrival, store powder at −20°C, solution at −80°C. Protect from light and avoid multiple freeze/thaw cycles.
sample preparation For detailed sample preparation 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).

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