DUB assay explorer panel

a panel of 4 deubiquitylating enzyme (DUB) activity assay reagents (25 μg each) prepared by total chemical synthesis

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Weight 0.01 kg
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350.00

Description

UbiQ-L03 is a panel of 4 deubiquitylating enzyme (DUB) activity assay reagents (25 μg each) prepared by total chemical synthesis.

  1. Ub-AMC (UbiQ-001) is a quenched fluorogenic DUB substrate based on the C-terminal derivatization of ubiquitin with 7-amido-4-methylcoumarin (AMC). Cleavage of the amide bond between Gly76 of ubiquitin and the AMC moiety results in an increase in fluorescence at λEx /λEx= 380/460 nm.
  2. Ub-Rh110Gly (UbiQ-002) is a quenched fluorogenic DUB substrate based on the C-terminal derivatization of ubiquitin with rhodamine110-Gly (Rh110Gly). Cleavage of the amide bond between Gly76 of ubiquitin and Rh110Gly results in an increase in fluorescence at λEx /λEx= 535/485 nm.
  3. TAMRA-Lys(Ub)-Gly (UbiQ-012) is a fluorescence polarization assay reagent based on a 5- carboxytetramethylrhodamine (TAMRA λEx /λEx= 530/580 nm) modified Lys-Gly sequence that is linked to ubiquitin via a native isopeptide bond with the lysine side-chain.
  4. Ub-aminoluciferin (UbiQ-036) is a DUB substrate based on the C-terminal derivatization of ubiquitin with 6-aminoluciferin (Luc). Upon cleavage by a DUB, the released 6-aminoluciferin functions as a substrate for luciferase, allowing detection of luminescence as a read-out for DUB-activity.

Additional information

Weight 0.01 kg
Applications

,

target

source

shipping

purity

storage

sample preparation

For detailed sample preparation see product sheet.

regulatory statement

aliquot size

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