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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

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

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

Sharp, P.M. & W.-H. Li. (1987) Trends Ecol. Evol. 2:328.
Ciechanover, A. et al. (1980 ) Proc. Natl. Acad. Sci. USA 77:1365.
Hershko, A. et al. (1980) Proc. Natl. Acad. Sci. USA 77:1783.
Greene, W. et al. (2012) PLoS Pathog. 8:e1002703.
Tong, X. et al. (2012) J. Biol. Chem. 287:25280.
Wei, W. et al. (2004) Nature 428:194.
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

Related Research Areas

Cell Structure and Signaling Molecules
Cell to Cell Adhesion Disassembly During EMT
HIF Transcription Factors
Negative Regulators of the Jak/STAT Pathway
Neurofibrillary Tangle Formation
NFkB Pathway
Notch Pathway
Parkinson"s Disease Related Molecules
Ubiquitin
Ubiquitin-related Molecules in the Akt Pathway
Wnt Intracellular Signaling

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.

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Stepwise multi-polyubiquitantion of p53 by the E6AP-E6 ubiquitin ligase complexAuthors: Y Masuda, Y Saeki, N Arai, H Kawai, I Kukimoto, K Tanaka, C MasutaniJ. Biol. Chem., 2019;0(0):.Species: HumanSample Types: Recombinant ProteinApplications: Bioassay
The pseudosubstrate inhibitor Acm1 inhibits the anaphase-promoting complex/cyclosome by combining high-affinity activator binding with disruption of Doc1/Apc10 functionAuthors: L Qin, A Mizrak, DSPSF Guimarães, HM Tambrin, DO Morgan, MC HallJ. Biol. Chem., 2019;0(0):.Species: YeastSample Types: ProteinApplications: Bioassay
Ubiquitination and dynactin regulate TMEPAI lysosomal traffickingAuthors: S Luo, L Jing, T Zhao, Y Li, Z Liu, A DiaoSci Rep, 2017;7(0):42668.Species: HumanSample Types: Cell LysatesApplications: Bioassay
Development of ?-Hairpin Peptides for the Measurement of SCF-Family E3 Ligase Activity in Vitro via Ornithine UbiquitinationAuthors: KM Houston, AT Melvin, GS Woss, EL Fayer, ML Waters, NL AllbrittonACS Omega, 2017;2(3):1198-1206.Species: HumanSample Types: ProteinApplications: Bioassay
Ufd2p synthesizes branched ubiquitin chains to promote the degradation of substrates modified with atypical chainsAuthors: C Liu, W Liu, Y Ye, W LiNat Commun, 2017;8(0):14274.Species: YeastSample Types: Cell LysatesApplications: Bioassay
RYBP stimulates PRC1 to shape chromatin-based communication between Polycomb repressive complexesElife, 2016;5(0):.Applications: Bioassay
Quantitative framework for ordered degradation of APC/C substrates.Authors: Lu D, Girard J, Li W, Mizrak A, Morgan DBMC Biol, 2015;13(0):96.Applications: Bioassay
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 JPLoS Pathog, 2012;8(12):e1003085.Species: N/ASample Types: Recombinant ProteinApplications: Bioassay
Ubiquitylation of terminal deoxynucleotidyltransferase inhibits its activity.Authors: Maezawa S, Fukushima R, Matsushita TPLoS ONE, 2012;7(7):e39511.Species: BovineSample Types: DNA
The APC/C subunit Mnd2/Apc15 promotes Cdc20 autoubiquitination and spindleassembly checkpoint inactivation.Authors: Foster S, Morgan DMol Cell, 2012;47(6):921-32.Species: N/ASample Types: ProteinApplications: Bioassay