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Recombinant Human FLAG (DYKDDDDK)-Ubiquitin Protein, CF

Catalog #: U-120
17 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
U-120-01M
R&D Systems Recombinant Proteins and Enzymes
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Citations (17)
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Recombinant Human FLAG (DYKDDDDK)-Ubiquitin Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.
Activity
Recombinant Human FLAG-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 an initial Recombinant Human FLAG-Ubiquitin concentration of 0.5-1 mM.
Source
E. coli-derived human Ubiquitin protein
Contains an N-terminal FLAG (DYKDDDDK) tag
Accession #
P0CG47
Predicted Molecular Mass
9.8 kDa

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Product Datasheets

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U-120

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-120

Formulation Lyophilized from a solution in deionized water.
Reconstitution Reconstitute at 10 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).

This fully functional N-terminal FLAG-tagged Ubiquitin protein allows for the convenient detection or affinity purification of ubiquitinated proteins in vitro. The epitope tag is a hydrophilic octapeptide (DYKDDDDK) derived from the sequence of the bacteriophage 7 gene-10 product. This tag is specifically recognized by Anti-FLAG antibodies M1, M2 or M5.

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 FLAG (DYKDDDDK)-Ubiquitin 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.

17 Citations: Showing 1 - 10
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  1. Proteasome-associated ubiquitin ligase relays target plant hormone-specific transcriptional activators
    Authors: Z Wang, B Orosa-Puen, M Nomoto, H Grey, T Potuschak, T Matsuura, IC Mori, Y Tada, P Genschik, SH Spoel
    Science Advances, 2022;8(42):eabn4466.
    Species: Complex Species Category
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. Identification of disease-linked hyperactivating mutations in UBE3A through large-scale functional variant analysis
    Authors: KP Weston, X Gao, J Zhao, KS Kim, SE Maloney, J Gotoff, S Parikh, YC Leu, KP Wu, M Shinawi, JP Steimel, JS Harrison, JJ Yi
    Nature Communications, 2021;12(1):6809.
    Species: Human
    Sample Types: Recombinant Proteins
    Applications: Bioassay
  3. Ubiquitylation of lipopolysaccharide by RNF213 during bacterial infection
    Authors: EG Otten, E Werner, A Crespillo-, KB Boyle, V Dharamdasa, C Pathe, B Santhanam, F Randow
    Nature, 2021;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Ubiquitylation
  4. Alternative splicing and allosteric regulation modulate the chromatin binding of UHRF1
    Authors: M Tauber, S Kreuz, A Lemak, P Mandal, Z Yerkesh, A Veluchamy, B Al-Gashgar, A Aljahani, LV Cortés-Med, D Azhibek, L Fan, MS Ong, S Duan, S Houliston, CH Arrowsmith, W Fischle
    Nucleic Acids Res., 2020;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Ubiquitination
  5. Ubiquitination of MAP1LC3B by pVHL is associated with autophagy and cell death in renal cell carcinoma
    Authors: HM Kang, KH Noh, TK Chang, D Park, HS Cho, JH Lim, CR Jung
    Cell Death Dis, 2019;10(4):279.
    Applications: Bioassay
  6. Ring finger protein 145 (RNF145) is a ubiquitin ligase for sterol-induced degradation of HMG-CoA reductase
    Authors: LY Jiang, W Jiang, N Tian, YN Xiong, J Liu, J Wei, KY Wu, J Luo, XJ Shi, BL Song
    J. Biol. Chem., 2018;0(0):.
    Applications: Bioassay
  7. Proteasome-associated HECT-type ubiquitin ligase activity is required for plant immunity
    Authors: JJ Furniss, H Grey, Z Wang, M Nomoto, L Jackson, Y Tada, SH Spoel
    PLoS Pathog., 2018;14(11):e1007447.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  8. Reciprocal Regulation between 53BP1 and the Anaphase-Promoting Complex/Cyclosome Is Required for Genomic Stability during Mitotic Stress
    Authors: TJ Kucharski, PE Minshall, M Moustafa-K, AS Turnell, JG Teodoro
    Cell Rep, 2017;18(8):1982-1995.
  9. Ufd2p synthesizes branched ubiquitin chains to promote the degradation of substrates modified with atypical chains
    Authors: C Liu, W Liu, Y Ye, W Li
    Nat Commun, 2017;8(0):14274.
    Species: Yeast
    Sample Types: Cell Lysates
    Applications: Bioassay
  10. The Ubiquitin Ligase CHIP Integrates Proteostasis and Aging by Regulation of Insulin Receptor Turnover
    Authors: R Tawo, W Pokrzywa, É Kevei, ME Akyuz, V Balaji, S Adrian, J Höhfeld, T Hoppe
    Cell, 2017;169(3):470-482.e13.
    Applications: Bioassay
  11. COP1 mediates dark-specific degradation of microtubule-associated protein WDL3 in regulating Arabidopsis hypocotyl elongation
    Authors: N Lian, X Liu, X Wang, Y Zhou, H Li, J Li, T Mao
    Proc. Natl. Acad. Sci. U.S.A., 2017;114(46):12321-12326.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  12. Ubiquitin- and ATP-dependent unfoldase activity of P97/VCP�NPLOC4�UFD1L is enhanced by a mutation that causes multisystem proteinopathy
    Authors: EE Blythe, KC Olson, V Chau, RJ Deshaies
    Proc. Natl. Acad. Sci. U.S.A., 2017;0(0):.
    Applications: Bioassay
  13. Hemi-methylated DNA regulates DNA methylation inheritance through allosteric activation of H3 ubiquitylation by UHRF1
    Elife, 2016;5(0):.
    Species: Human
    Sample Types: Protein
    Applications: Ubiquitination
  14. Dimeric Ube2g2 simultaneously engages donor and acceptor ubiquitins to form Lys48-linked ubiquitin chains.
    Authors: Liu, Weixiao, Shang, Yonglian, Zeng, Yan, Liu, Chao, Li, Yanchang, Zhai, Linhui, Wang, Pan, Lou, Jizhong, Xu, Ping, Ye, Yihong, Li, Wei
    EMBO J, 2014;33(1):46-61.
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Bioassay
  15. The Rad6/18 ubiquitin complex interacts with the Epstein-Barr virus deubiquitinating enzyme, BPLF1, and contributes to virus infectivity.
    Authors: Kumar, Ravindra, Whitehurst, Christop, Pagano, Joseph S
    J Virol, 2014;88(11):6411-22.
    Species: N/A
    Sample Types: Protein
    Applications: Bioassay
  16. Cytosolic quality control of mislocalized proteins requires RNF126 recruitment to Bag6.
    Authors: Rodrigo-Brenni M, Gutierrez E, Hegde R
    Mol Cell, 2014;55(2):227-37.
    Applications: Bioassay
  17. An unusual two-step control of CPEB destruction by Pin1.
    Authors: Nechama, Morris, Lin, Chien-Li, Richter, Joel D
    Mol Cell Biol, 2013;33(1):48-58.
    Species: Xenopus
    Sample Types: In Vivo
    Applications: In Vivo

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Reconstitution Buffers

Reconstitution Buffer 1 (PBS)

RB01
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