Ub-Ub(56-69)-FP K63 linked

a class II ubiquitinated peptide fluorescence polarization assay reagent based on the peptide sequence 56 – 69 of ubiquitin

productsheet

code UbiQ-049 Category
product UbiQ-049 Category

Additional information

Weight 0.005 kg
aliquot size

Applications

,

target

source

shipping

purity

molecular weight

storage

sample preparation

Prepare a 2 mM stock in DMSO (21 mg/mL). Dilute the 2 mM DMSO stock 20x in milliQ affording a 100 μM stock which can be aliquoted and stored. For assays this 100 μM stock can be diluted for example 1000x in buffer affording a final assay solution of 100 nM, the DMSO concentration during the assay is now 0.01 vol%. The concentration can be verified by comparing the fluorescence intensity with that of a known concentration of TAMRA.

regulatory statement

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300.00

Description

UbiQ-049 is a class II ubiquitinated peptide fluorescence polarization assay reagent based on the peptide sequence 56 – 69 of ubiquitin.
The peptide is modified on the N-terminus with a 5-carboxytetramethylrhodamine and conjugated at Lys63 to Ub via a native isopeptide bond.
See reference 5 in the product sheet (open access) for full experimental and analytical details.

Additional information

Weight 0.005 kg
aliquot size

Applications

,

target

source

shipping

purity

molecular weight

storage

sample preparation

Prepare a 2 mM stock in DMSO (21 mg/mL). Dilute the 2 mM DMSO stock 20x in milliQ affording a 100 μM stock which can be aliquoted and stored. For assays this 100 μM stock can be diluted for example 1000x in buffer affording a final assay solution of 100 nM, the DMSO concentration during the assay is now 0.01 vol%. The concentration can be verified by comparing the fluorescence intensity with that of a known concentration of TAMRA.

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

Faesen, A.C., et al. The Differential Modulation of USP Activity by Internal Regulatory Domains, Interactors and Eight Ubiquitin Chain Types. Chem. Biol. 18, 1550-1561 (2011).
http://www.ncbi.nlm.nih.gov/pubmed/22195557