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Recombinant Human Ubiquitin N-Terminal Biotin Protein, CF

Catalog #: UB-560
6 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
UB-560-050
R&D Systems Recombinant Proteins and Enzymes
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Citations (6)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Ubiquitin N-Terminal Biotin Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.
Activity
Biotinylated Recombinant Human Ubiquitin can be conjugated to substrate proteins via the subsequent actions of a Ubiquitin-activating (E1) enzyme, a Ubiquitin-conjugating (E2) enzyme, and a Ubiquitin ligase (E3). Reaction conditions will need to be optimized for each specific application. We recommend using Biotinylated Recombinant Human Ubiquitin in conjunction with native Ubiquitin at a combined concentration of 10-50 μM with a 1:2 to 1:20 ratio of Biotinylated Recombinant Human Ubiquitin:native Ubiquitin. The resulting poly-Ubiquitin chains can be visualized/quantitated with avidin-linked detection reagents.
Source
E. coli-derived human Ubiquitin protein
Gln2 - Gly76 with a single N‑terminal biotin
Accession #
P0CG47.1
Predicted Molecular Mass
9 kDa

Product Datasheets

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UB-560

Product Datasheet
COA
SDS

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.

UB-560

Formulation Supplied as a solution in 10 mM HEPES pH 8.0
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
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).

Produced via a proprietary process resulting in a single Biotin modification exclusively on the N-terminus of Ubiquitin. This site-specific modification results in an Ubiquitin that is fully functional at the C-terminus, and with the full compliment of reactive lysines to allow for poly-Ubiquitin chain incorporation. Use of Avidin-linked reagents allows for high efficiency capture and/or detection sensitivity. Ideal as an alternative To radio-labeled Ubiquitin

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.
Entrez Gene IDs
7314 (Human); 298693 (Rat)
Alternate Names
RPS27A; UBA52; UBB ubiquitin B; UBB; UBC; Ubiquitin

Citations for Recombinant Human Ubiquitin N-Terminal Biotin 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.

6 Citations: Showing 1 - 6
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  1. Substitution of PINK1 Gly411 modulates substrate receptivity and turnover
    Authors: FC Fiesel, D Fri?ová, CS Hayes, MA Coban, R Hudec, JM Bredenberg, BJ Broadway, BN Markham, T Yan, PK Boneski, G Fiorino, JO Watzlawik, X Hou, AM McCarty, LJ Lewis-Tuff, J Zhong, BJ Madden, A Ordureau, H An, A Puschmann, ZK Wszolek, OA Ross, JW Harper, TR Caulfield, W Springer
    Autophagy, 2022-12-05;0(0):1-22.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  2. Substitution of PINK1 Gly411 modulates substrate receptivity and turnover
    Authors: FC Fiesel, D Fri?ová, CS Hayes, MA Coban, R Hudec, JM Bredenberg, BJ Broadway, BN Markham, T Yan, PK Boneski, G Fiorino, JO Watzlawik, X Hou, AM McCarty, LJ Lewis-Tuff, J Zhong, BJ Madden, A Ordureau, H An, A Puschmann, ZK Wszolek, OA Ross, JW Harper, TR Caulfield, W Springer
    Autophagy, 2022;0(0):1-22.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  3. Age- and disease-dependent increase of the mitophagy marker phospho-ubiquitin in normal aging and Lewy body disease
    Authors: X Hou, FC Fiesel, D Truban, M Castanedes, WL Lin, AI Soto, P Tacik, LG Rousseau, NN Diehl, MG Heckman, O Lorenzo-Be, I Ferrer, JM Arbelo, JC Steele, MJ Farrer, M Cornejo-Ol, L Torres, IF Mata, NR Graff-Radf, ZK Wszolek, OA Ross, ME Murray, DW Dickson, W Springer
    Autophagy, 2018-07-28;14(8):1404-1418.
    Applications: Bioassay
  4. The PINK1 p.I368N mutation affects protein stability and ubiquitin kinase activity
    Authors: M Ando, FC Fiesel, R Hudec, TR Caulfield, K Ogaki, P Górka-Skoc, D Koziorowsk, A Friedman, L Chen, VL Dawson, TM Dawson, G Bu, OA Ross, ZK Wszolek, W Springer
    Mol Neurodegener, 2017-04-24;12(1):32.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  5. Heterozygous PINK1 p.G411S increases risk of Parkinson's disease via a dominant-negative mechanism
    Authors: Wolfdieter Springer
    Brain, 2016-11-02;0(0):.
    Applications: Bioassay
  6. The E3 ligase HOIP specifies linear ubiquitin chain assembly through its RING-IBR-RING domain and the unique LDD extension.
    EMBO J., 2012-08-03;31(19):3833-44.
    Applications: Ubiquitination

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