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Recombinant Human UbcH5c/UBE2D3 - Ubiquitin Charged Protein

Catalog #: E2-802
2 Citations Datasheet / COA / SDS

Discontinued Product

E2-802 has been discontinued.
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Recombinant Human UbcH5c/UBE2D3 - Ubiquitin Charged Protein Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain
Activity

Pre-charged UBE2D3 may be added directly to in vitro reactions containing E3 Ubiquitin ligases and ligase substrates (if applicable)—no ATP, E1 enzyme, or extra Ubiquitin are required.  Reaction conditions will need to be optimized for each specific application.  Note: Reducing agents including dithiothreitol (DTT) or mercaptoethanol ( beta ME) may cause unintended thiolytic release of Ubiquitin from the complex—care must be taken if these compounds are present in buffers. We suggest using a thioester friendly reductant such as TCEP if possible.

Source
E. coli-derived human UbcH5c/UBE2D3 protein
Accession # P61077, P0CG47
Accession #
P61077, P0CG47
Predicted Molecular Mass

25 kDa (17 kDa UBE2D3, 8.6 kDa Ubiquitin)

Product Datasheets

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E2-802

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.

E2-802

Formulation

Approximately 0.43 mg/ml (25 μM) with respect to UBE2D3.  Formulated in 50 mM HEPES pH 7.5, 50 mM NaCl, 1 mM TCEP
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -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: UbcH5c/UBE2D3

Ubiquitin­-conjugating Enzyme H5c (UbcH5c), also known as Ubiquitin­-conjugating Enzyme E2D 3 (UBE2D3), is a member of the yeast Ubc4/5 family of Ubiquitin­conjugating (E2) enzymes. Human UbcH5c/UBE2D3 has a predicted molecular weight of 17 kDa and shares 88% and 89% amino acid sequence identity with the related family members, UbcH5a and UbcH5b, respectively.  In combination with Ubiquitin ligases (E3s) such as CHIP, UbcH5c/UBE2D3 mediates the ubiquitination and subsequent degradation of several regulatory proteins.  For instance, UbcH5c/UBE2D3 is involved in the polyubiquitination and proteasome­mediated degradation of the Nuclear Factor kappaB (NF­ kappa B) inhibitor, I kappa B­ alpha, and is implicated in NF­ kappa B­-dependent inflammation.  UbcH5c/UBE2D3 also mediates the ubiquitination of Histone H2A and PCNA, suggesting that it functions during transcriptional regulation, DNA replication, and DNA damage responses.

This product is an enzymatically generated UbcH5c/UBE2D3-Ubiquitin thioester complex that has been purified to remove E1 Ubiquitin Activating enzyme, uncharged UbcH5c/UBE2D3, free Ubiquitin, and Mg2+-ATP.  The product provides a convenient starting material for use in single-turnover "Ubiquitin Discharge Assays," eliminating the need to either inhibit the E1 Ubiquitin Activating enzyme with potentially confounding chemical treatments or remove ATP via enzyme additions.

References
  1. Das, R. et al. (2013) EMBO J. 32: 2504
  2. Koyano, F. et al. (2014) Nature 510: 162
  3. Liew, C.W. et al. (2010) Biochem. J. 431: 23
  4. Petroski, M.D. & Deshaies, R.J. (2005) Cell 123: 1107
  5. Saha, A. et al. (2011) Mol. Cell 42: 75
Entrez Gene IDs
7323 (Human); 66105 (Mouse); 81920 (Rat)
Alternate Names
E2(17)KB 3; E2(17)KB3; EC 6.3.2.19; MGC43926; MGC5416; UBC4/5; UBC5C; UbcH5c; UBE2D3; Ubiquitin carrier protein D3; ubiquitin-conjugating enzyme E2 D3; Ubiquitin-conjugating enzyme E2(17)KB 3; Ubiquitin-conjugating enzyme E2-17 kDa 3; ubiquitin-conjugating enzyme E2D 3 (homologous to yeast UBC4/5); ubiquitin-conjugating enzyme E2D 3 (UBC4/5 homolog, yeast); Ubiquitin-protein ligase D3

Citations for Recombinant Human UbcH5c/UBE2D3 - Ubiquitin Charged Protein

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. Regulated interaction of ID2 with the anaphase-promoting complex links progression through mitosis with reactivation of cell-type-specific transcription
    Authors: SB Lee, L Garofano, A Ko, F D'Angelo, B Frangaj, D Sommer, Q Gan, K Kim, T Cardozo, A Iavarone, A Lasorella
    Nature Communications, 2022-04-19;13(1):2089.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  2. Targeting a helix-in-groove interaction between E1 and E2 blocks ubiquitin transfer
    Authors: AM Cathcart, GH Bird, TE Wales, HD Herce, EP Harvey, ZJ Hauseman, CE Newman, U Adhikary, MS Prew, T Oo, S Lee, JR Engen, LD Walensky
    Nat. Chem. Biol., 2020-08-17;0(0):.
    Species: Human, N/A
    Sample Types: Peptide, Protein
    Applications: Bioassay

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