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Recombinant Human Ubiquitin Biotin Protein, CF

Catalog #: UB-570
6 Citations Datasheet / COA / SDS

Discontinued Product

UB-570 has been discontinued.
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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 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 100-500 μM with a 1:1 to 1:5 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
Accession #
P0CG47
Predicted Molecular Mass
8.6 kDa (unlabeled), extent of biotinylation varies by lot

Product Datasheets

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UB-570

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-570

Formulation X mg/ml in 10 mM HEPES pH 7.5
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 24 months from date of receipt, -20 to -70 °C as supplied.
  • 12 months, -20 to -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).

Ubiquitin modified with Biotin via primary amine coupling results in multiple biotinylated Ubiquitin species modified at the N-terminus, as well as lysine residues. Although having a fully functional C-terminus, lysine modification may limit the ability of this reagent to propagate poly-Ubiquitin chains. Biotinylated Ubiquitin can be detected using avidin-linked reagents.

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 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. Parkin Precipitates on Mitochondria via Aggregation and Autoubiquitination
    Authors: Ardah, MT;Radwan, N;Khan, E;Kitada, T;Haque, ME;
    International journal of molecular sciences
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. Antibody toolkit reveals N-terminally ubiquitinated substrates of UBE2W
    Authors: CW Davies, SE Vidal, L Phu, J Sudhamsu, TB Hinkle, S Chan Rosen, FR Schumacher, YJ Zeng, C Schwerdtfe, AS Peterson, JR Lill, CM Rose, AS Shaw, IE Wertz, DS Kirkpatric, JT Koerber
    Nature Communications, 2021-07-29;12(1):4608.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  3. Robust high-throughput assays to assess discrete steps in ubiquitination and related cascades
    Authors: G Fenteany, P Gaur, G Sharma, L Pintér, E Kiss, L Haracska
    BMC Mol Cell Biol, 2020-03-30;21(1):21.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. PI5P4K? functions in DTX1-mediated Notch signaling
    Authors: L Zheng, SD Conner
    Proc. Natl. Acad. Sci. U.S.A., 2018-02-12;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  5. Opposing effects of cancer-type-specific SPOP mutants on BET protein degradation and sensitivity to BET inhibitors
    Authors: H Janouskova, G El Tekle, E Bellini, ND Udeshi, A Rinaldi, A Ulbricht, T Bernasocch, G Civenni, M Losa, T Svinkina, CM Bielski, GV Kryukov, L Cascione, S Napoli, RI Enchev, DG Mutch, ME Carney, A Berchuck, BJN Winterhoff, RR Broaddus, P Schraml, H Moch, F Bertoni, CV Catapano, M Peter, SA Carr, LA Garraway, PJ Wild, JP Theurillat
    Nat. Med., 2017-08-14;23(9):1046-1054.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  6. Ubiquitin Modification by the E3 Ligase/ADP-Ribosyltransferase Dtx3L/Parp9
    Authors: CS Yang, K Jividen, A Spencer, N Dworak, L Ni, LT Oostdyk, M Chatterjee, B Ku?mider, B Reon, M Parlak, V Gorbunova, T Abbas, E Jeffery, NE Sherman, BM Paschal
    Mol. Cell, 2017-05-18;66(4):503-516.e5.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay

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