Recombinant Human Ubiquitin Mutant with K63 only Protein, CF

 

Discontinued Product

UM-K630 has been discontinued.
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Citations (5)
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Recombinant Human Ubiquitin Mutant with K63 only Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.
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 linkage. 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 an initial Recombinant Human Ubiquitin K63-only concentration of 0.2-1 mM.
Source
E. coli-derived human Ubiquitin protein
Accession #
Predicted Molecular Mass
8.7 kDa

Product Datasheets

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UM-K630

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

UM-K630

Formulation Lyophilized from a solution in deionized water.
Reconstitution

Reconstitute at 2 mg/mL in an aqueous solution.

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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

This Ubiquitin mutant contains only a single lysine, K63, with all other lysines mutated to arginine. This mutation renders Ubiquitin able to form poly-Ubiquitin chains with other Ubiquitin molecules only via the K63 lysine.

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.
  8. Arnason T., et al. (1994) Mol. Cell. Biol. 14:7876-7883.
  9. Spence J., et al. (1995) Mol. Cell. Biol. 15:1265-1273.  
Entrez Gene IDs
7314 (Human); 298693 (Rat)
Alternate Names
RPS27A; UBA52; UBB ubiquitin B; UBB; UBC; Ubiquitin

Citations for Recombinant Human Ubiquitin Mutant with K63 only Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Multi-tiered actions of Legionella effectors to modulate host Rab10 dynamics
    Authors: Kubori, T;Arasaki, K;Oide, H;Kitao, T;Nagai, H;
    eLife
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. A distinct mammalian disome collision interface harbors K63-linked polyubiquitination of uS10 to trigger hRQT-mediated subunit dissociation
    Authors: M Narita, T Denk, Y Matsuo, T Sugiyama, C Kikuguchi, S Ito, N Sato, T Suzuki, S Hashimoto, I Machová, P Tesina, R Beckmann, T Inada
    Nature Communications, 2022-10-27;13(1):6411.
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Bioassay
  3. Site-specific ubiquitination of MLKL targets it to endosomes and targets Listeria and Yersinia to the lysosomes
    Authors: S Yoon, K Bogdanov, D Wallach
    Cell Death and Differentiation, 2022-01-09;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  4. Novel polyubiquitin imaging system, PolyUb-FC, reveals that K33-linked polyubiquitin is recruited by SQSTM1/p62
    Authors: Y Nibe, S Oshima, M Kobayashi, C Maeyashiki, Y Matsuzawa, K Otsubo, H Matsuda, E Aonuma, Y Nemoto, T Nagaishi, R Okamoto, K Tsuchiya, T Nakamura, S Nakada, M Watanabe
    Autophagy, 2018-01-24;0(0):1-43.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  5. p47 negatively regulates IKK activation by inducing the lysosomal degradation of polyubiquitinated NEMO.
    Authors: Shibata Y, Oyama M, Kozuka-Hata H, Han X, Tanaka Y, Gohda J, Inoue J
    Nat Commun, 2012-01-01;3(0):1061.
    Applications: Binding Assay

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Recombinant Human Ubiquitin Mutant with K63 only Protein, CF
By Anonymous on 10/26/2017
Application: Enzymatic activity in vitro