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Ubiquitin-AMC

High purity industry standard substrate for deubiquitinylating enzymes (DUBs).
 
BML-SE211-0025 25 µg 192.00 USD
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Ubiquitin-AMC is a fluorogenic substrate for a wide range of deubiquitinylating enzymes (DUBs), including ubiquitin C-terminal hydrolases (UCHs) and ubiquitin specific proteases (USPs). It is a particularly useful reagent for the study of deubiquitinylating activity where detection sensitivity or continuous monitoring of activity is essential. Ubiquitin-AMC has been shown to be a sensitive substrate for UCH-L3 (Km = 0.039μM) (Prod. No. BML-UW9745) and for Isopeptidase-T (Km = 0.17-1.4μM) (Prod. No. BML-UW9690), and is particularly useful for studying deubiquitinylating activity where detection sensitivity or continuous monitoring of activity is essential.

Product Specification

Source:Produced in E.coli.
 
UniProt ID:P0CG47 (UBB), P0CG48 (UBC), P62979 (RPS27A), P62987 (UBA52)
 
Formulation:In DMSO
 
Purity:≥95% (HPLC)
 
Purity Detail:Please note: considerable efforts are taken to ensure that ubiquitin-AMC is homogeneous and not contaminated with ubiquitin and ubiquitin deletions, which are common in material obtained from other sources.  The chromatogram below demonstrates the homogeneity of BML-SE211.
 
Application Notes:Can be used for the following applications:
1. Substrate for deubiquitinylating enzyme activity assays.
2. Identification/confirmation of enzyme deubiquitinylation activity.
3. Investigation of deconjugating enzyme substrate specificity in comparison with alternative UBL substrates.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Handling:Avoid freeze/thaw cycles.
 
bml-se211

Figure:  HPLC analysis. Comparison of Prod. No. BML-SE-211Prod. No. BML-SE-211) (A) with contaminated alternatively sourced material (B).

 

bml-se211 b
Figure: DUB Assay: 50nM UCLH1, His6-tagged (UW9740); 500nM Ubiquitin-AMC; 50mM HEPES pH7.8, 0.5mM EDTA, 1mM DTT.
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bml-se211 bml-se211 b

Product Literature References

Ubiquitin Carboxy-Terminal HydrolaseL3 Correlates with Human Sperm Count, Motility and Fertilization: M. Wang, et al.; PLoS One 11, e0165198 (2016), Application(s): Determination of UCH enzymatic activity of spermatozoa , Abstract; Full Text
Substrate profiling of deubiquitin hydrolases with a positional scanning library and mass spectrometry: D. E. Mason et al.; Biochemistry 43, 6535 (2004), Abstract;
Kinetic and mechanistic studies on the hydrolysis of ubiquitin C-terminal 7-amido-4-methylcoumarin by deubiquitinating enzymes: L. C. Dang et al.; Biochemistry 37, 1868 (1998), Abstract;

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