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Recombinant Human HA-Ubiquitin Aldehyde Protein, CF

Catalog #: U-211
2 Citations Datasheet / COA / SDS

Discontinued Product

U-211 has been discontinued.
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Recombinant Human HA-Ubiquitin Aldehyde Protein, CF Summary

Product Specifications

Purity
>95%, by HPLC
Activity
Add Recombinant Human HA-Ubiquitin Aldehyde to in vitro assays to inhibit deubiquitinating enzymes. The HA-tag allows for detection and purification of deubiquitinating enzyme activity. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human HA-Ubiquitin Aldehyde concentration of 2-5 μM.
Source
E. coli-derived human Ubiquitin protein
Contains an N-terminal HA (YPYDVPDYA) tag and a mixture of derivatized and underivatized C-terminal Aldehyde, quantity is by derivatized content.
Accession #
P0CG47
Predicted Molecular Mass
9.8 kDa

Product Datasheets

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U-211

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.

U-211

Formulation Lyophilized from a solution in MES.
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.
  • 3 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).

Ubiquitin-aldehyde is a potent and specific inhibitor of most deubiquitinating enzymes (DUBs) such as Ubiquitin C-terminal hydrolases (UCHs) and Ubiquitin-specific proteases (USPs). It prevents the hydrolysis of poly-Ubiquitin chains on substrate proteins in vitro and thus enhances poly-Ubiquitin chain accumulation. This tagged version contains an N-terminal HA peptide sequence (YPYDVPDYA) derived from the influenza Hemagglutinin protein. This epitope allows for the sensitive identification or purification of DUBs since it is specifically recognizedby Anti-HA antibodies and/or Anti-HA-agarose.

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. Borodovsky A., et al. (2001) EMBO J. 20: 5187-5196.
  9. Callis J. and Ling R. (2005) Meth. Enzymol. 399:51-64.
  10. Catic A., et al. (2007) PLoS One 2:e679.
  11. Galardy P., et al. (2005) Methods Enz. 399:120-131.
  12. Gredmark S., et al. (2007) J.Virol. 81:10300-10309.
  13. Hemelaar J., et al. (2004) Mol. Cel. Biol. 24:84-95.
  14. Love K.R., et al. (2007) Nat. Chem. Biol. 3:697-705.
  15. Ovaa H., et al. (2005) Methods Enz. 399:468-478.
Entrez Gene IDs
7314 (Human); 298693 (Rat)
Alternate Names
RPS27A; UBA52; UBB ubiquitin B; UBB; UBC; Ubiquitin

Citations for Recombinant Human HA-Ubiquitin Aldehyde 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.

2 Citations: Showing 1 - 2
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  1. Mutual regulation between OGT and XIAP to control colon cancer cell growth and invasion
    Authors: HG Seo, HB Kim, JY Yoon, TH Kweon, YS Park, J Kang, J Jung, S Son, EC Yi, TH Lee, WH Yang, JW Cho
    Cell Death Dis, 2020-09-29;11(9):815.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. PCNA-Dependent Cleavage and Degradation of SDE2 Regulates Response to Replication Stress
    PLoS Genet, 2016-12-01;12(12):e1006465.
    Species: Human
    Sample Types: Whole Cells
    Applications: Enzyme Assay

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