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Recombinant Human Ubiquitin w/K63 only N-Term Biotin, CF

R&D Systems, part of Bio-Techne | Catalog # UB-K630

R&D Systems, part of Bio-Techne
Discontinued Product
UB-K630 has been discontinued. View all Ubiquitin products.

Key Product Details

Source

E. coli

Accession #

Conjugate

Biotin

Applications

Enzyme Activity

Product Specifications

Source

E. coli-derived human Ubiquitin protein

Purity

>95%, by PAGE.

Activity

The lysine residue utilized for Ubiquitin chain formation is functionally important. Single lysine Ubiquitin mutants are ideal for investigating biological processes involving specific Ubiquitin chain linkages. Biotinylated Recombinant Human Ubiquitin K63-only allows for the formation of only K63-linked Ubiquitin chains. Reaction conditions will need to be optimized for each specific application. We recommend using Biotinylated Recombinant Human Ubiquitin K63-only in conjunction with native Ubiquitin at a combined concentration of 10-50 μM with a 1:2 to 1:20 ratio of mutant Ubiquitin:native Ubiquitin. The resulting poly-Ubiquitin chains can be visualized/quantitated with avidin-linked detection reagents.

Formulation, Preparation and Storage

UB-K630
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Ubiquitin

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).

Produced via a proprietary process resulting in a single Biotin modification exclusively on the N-terminus of Ubiquitin. This site-specific modification results in a Ubiquitin that is fully functional at the C-terminus, and with the full compliment of reactive lysines to allow for poly-Ubiquitin chain incorporation. Detection with Avidin-linked reagents allows for a higher efficiency and detection sensitivity than with Anti-Ubiquitin antibodies. Ideal as an alternative Toradio-labeled Ubiquitin. This Ubiquitin mutant contains only a single lysine (K63) with all other lysines mutated to arginine, and is able to form poly-Ubiquitin chains with other Ubiquitin molecules via the K63 lysine only.

References

  1. Sharp, P.M. & W.-H. Li. (1987) Trends Ecol. Evol. 2:328.
  2. Ciechanover, A. et al. (1980 ) Proc. Natl. Acad. Sci. USA 77:1365.
  3. Hershko, A. et al. (1980) Proc. Natl. Acad. Sci. USA 77:1783.
  4. Greene, W. et al. (2012) PLoS Pathog. 8:e1002703.
  5. Tong, X. et al. (2012) J. Biol. Chem. 287:25280.
  6. Wei, W. et al. (2004) Nature 428:194.
  7. Wertz, I.E. et al. (2004) Nature 430:694.

Alternate Names

UBB

Entrez Gene IDs

7314 (Human); 298693 (Rat)

Gene Symbol

UBB

UniProt

Additional Ubiquitin Products

Product Documents for Recombinant Human Ubiquitin w/K63 only N-Term Biotin, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human Ubiquitin w/K63 only N-Term Biotin, CF

For research use only

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