Reactivity | HuSpecies Glossary |
Applications | Enzyme Activity |
Format | Carrier-Free |
Details of Functionality | Recombinant Human Ubiquitin-AMC is a fluorogenic substrate for some Ubiquitin-specific isopeptidases. Release of AMC fluorescence can be monitored with an excitation wavelength of 345 nm and an emission wavelength of 445 nm. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human Ubiquitin-AMC concentration of 0.1-1 μM. |
Source | E. coli-derived human Ubiquitin protein |
Accession # | |
Protein/Peptide Type | Recombinant Proteins |
Gene | UBB |
Dilutions |
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Theoretical MW | 8.7 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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Publications |
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Storage | Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Buffer | Supplied as a solution in DMSO. |
Ubiquitin is a 76 amino acid (aa) protein that is ubiquitously expressed in all eukaryotic organisms. Ubiquitin is highly conserved with 96% aa sequence identity shared between human and yeast Ubiquitin, and 100% aa sequence identity shared between human and mouse Ubiquitin (1). In mammals, four Ubiquitin genes encode for two Ubiquitin-ribosomal fusion proteins and two poly-Ubiquitin proteins. Cleavage of the Ubiquitin precursors by deubiquitinating enzymes gives rise to identical Ubiquitin monomers each with a predicted molecular weight of 8.6 kDa. Conjugation of Ubiquitin to target proteins involves the formation of an isopeptide bond between the C-terminal glycine residue of Ubiquitin and a lysine residue in the target protein. This process of conjugation, referred to as ubiquitination or ubiquitylation, is a multi-step process that requires three enzymes: a Ubiquitin-activating (E1) enzyme, a Ubiquitin-conjugating (E2) enzyme, and a Ubiquitin ligase (E3). Ubiquitination is classically recognized as a mechanism to target proteins for degradation and as a result, Ubiquitin was originally named ATP-dependent Proteolysis Factor 1 (APF-1) (2,3). In addition to protein degradation, ubiquitination has been shown to mediate a variety of biological processes such as signal transduction, endocytosis, and post-endocytic sorting (4-7).
This fluorogenic substrate for Ubiquitin hydrolases is based on the C-terminus derivatization of Ubiquitin with 7-amino-4-methylcoumarin (AMC). Ubiquitin-AMC is an exquisitely sensitive substrate for UCHL3 (Km = 0.039 μM) and for Isopeptidase-T (Km = 0.17 - 1.4 μM). Ubiquitin-AMC is useful for studying Ubiquitin hydrolases when detection sensitivity or continuous monitoring of activity is essential.
There's an autophagy for that! By Christina Towers, PhDA critical mechanism that cells use to generate nutrients and fuel metabolism is through a process called autophagy. This process is complex and involves over 20 different proteins, most of which are highly conserved acro... Read full blog post. |
Article Review: Glucose-induced transcriptional regulation in cancer Epigenetic mechanisms have been implicated in many physiological and pathophysiological processes. Among these, histone modifications including methylation, phosphorylation, acetylation and ubiquitination, significantly modify gene expression. In c... Read full blog post. |
PINK1: All work and no fun The protein PINK1 is a mitochondrial-located serine/threonine kinase (PTK) that maintains organelle function and integrity. It not only protects organelles from cellular stress, but it also uses the selective auto-phagocytosis process for cleaning and... Read full blog post. |
Ubiquitin-Mediated Degradation of Cellular Proteins: The Kiss of Death Ubiquitin is an abundant and essential cellular 9-kd protein that is conserved across evolution from yeast to humans. Ubiquitin is used by cells as a covalent modifier of other proteins both to activate their function and to target them for degradatio... Read full blog post. |
Using Ubiquitin Antibodies in Various Disease Research Ubiquitin is a small, highly conserved protein which plays an important role in protein breakdown, covalently bonding to proteins to mark them for proteolytic degradation in a process called ubiquitination. Ubiquitin also binds to inclusion bodies (ac... Read full blog post. |
The Heat is On: Heat Shock Proteins and the Link to Cancer Novus Biologicals offers an extensive antibody catalog targeting heat shock proteins (HSPs). A large protein group covering a number of families, the HSPs are functionally related by their dramatic upregulation in response to stress. Stress triggers m... Read full blog post. |
The Latest Research on IBR-type E3 Ubiquitin Ligases E3 ubiquitin ligases are standards in most antibody catalogs. These proteins are essential to the process of ubiquitination, which is expressed in protein pathways throughout the body and is often linked to disease states. It is widely used as a bioma... Read full blog post. |
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