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Recombinant Human Methylated Ubiquitin Protein, CF

Catalog #: U-501
10 Citations Datasheet / COA / SDS
 

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U-501 has been discontinued.
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Citations (10)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Methylated Ubiquitin Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain
Activity
Recombinant Human Methylated 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). Recombinant Human Methylated Ubiquitin is unable to form chains, making it ideal for use as a negative control for chain formation or to confirm multi-mono-ubiquitination. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human Methylated Ubiquitin concentration of 0.2-1 mM.
Source
E. coli-derived human Ubiquitin protein
Accession #
P0CG47
Predicted Molecular Mass
8.7 kDa

Product Datasheets

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U-501

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-501

Formulation Lyophilized from a solution in deionized water.
Reconstitution Reconstitute at 10 mg/mL in an aqueous solution.
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.
  • 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).

Reductive methylation of lysine residues renders Ubiquitin unable to form poly-Ubiquitin chains via lysine linkages with other Ubiquitin molecules. Methylated Ub can form a Ubiquitin-activating (E1) enzyme-catalyzed active thioester at the C-terminus allowing the molecule to be transferred to the lysines of substrate proteins (which can be mono-ubiquitinated). This is ideal for the reduction in poly-Ubiquitin chain length and rates of Ubiquitin conjugation.

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 Methylated 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.

10 Citations: Showing 1 - 10
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  1. The pseudosubstrate inhibitor Acm1 inhibits the anaphase-promoting complex/cyclosome by combining high-affinity activator binding with disruption of Doc1/Apc10 function
    Authors: L Qin, A Mizrak, DSPSF Guimarães, HM Tambrin, DO Morgan, MC Hall
    J. Biol. Chem., 2019-09-27;0(0):.
    Species: Yeast
    Sample Types: Protein
    Applications: Bioassay
  2. Stepwise multi-polyubiquitantion of p53 by the E6AP-E6 ubiquitin ligase complex
    Authors: Y Masuda, Y Saeki, N Arai, H Kawai, I Kukimoto, K Tanaka, C Masutani
    J. Biol. Chem., 2019-09-06;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  3. Development of ?-Hairpin Peptides for the Measurement of SCF-Family E3 Ligase Activity in Vitro via Ornithine Ubiquitination
    Authors: KM Houston, AT Melvin, GS Woss, EL Fayer, ML Waters, NL Allbritton
    ACS Omega, 2017-03-29;2(3):1198-1206.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  4. Ubiquitination and dynactin regulate TMEPAI lysosomal trafficking
    Authors: S Luo, L Jing, T Zhao, Y Li, Z Liu, A Diao
    Sci Rep, 2017-02-20;7(0):42668.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  5. 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, 2017-02-06;8(0):14274.
    Species: Yeast
    Sample Types: Cell Lysates
    Applications: Bioassay
  6. RYBP stimulates PRC1 to shape chromatin-based communication between Polycomb repressive complexes
    Elife, 2016-10-05;5(0):.
    Applications: Bioassay
  7. Quantitative framework for ordered degradation of APC/C substrates.
    Authors: Lu D, Girard J, Li W, Mizrak A, Morgan D
    BMC Biol, 2015-11-16;13(0):96.
    Applications: Bioassay
  8. Inhibition of a NEDD8 Cascade Restores Restriction of HIV by APOBEC3G.
    Authors: Stanley D, Bartholomeeusen K, Crosby D, Kim D, Kwon E, Yen L, Cartozo N, Li M, Jager S, Mason-Herr J, Hayashi F, Yokoyama S, Krogan N, Harris R, Peterlin B, Gross J
    PLoS Pathog, 2012-12-27;8(12):e1003085.
    Applications: Bioassay
  9. The APC/C subunit Mnd2/Apc15 promotes Cdc20 autoubiquitination and spindle assembly checkpoint inactivation.
    Authors: Foster S, Morgan D
    Mol Cell, 2012-08-30;47(6):921-32.
    Applications: Bioassay
  10. Ubiquitylation of terminal deoxynucleotidyltransferase inhibits its activity.
    Authors: Maezawa S, Fukushima R, Matsushita T
    PLoS ONE, 2012-07-11;7(7):e39511.
    Species: Bovine
    Sample Types: DNA

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