Recombinant Human Di-Ub (K48-linked) FRET TAMRA Pos1, CF

 

Discontinued Product

UF-210 has been discontinued.
View all Di-Ubiquitin products.
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Recombinant Human Di-Ub (K48-linked) FRET TAMRA Pos1, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain
Activity
Di-Ub (K48-linked) FRET TAMRA Pos1 is ideal for use as a substrate for Ubiquitin-specific hydrolases with activity toward K48 linkages. We recommend an assay buffer consisting of 50 mM Hepes pH 7.5, 150 mM NaCl, 2 mM DTT. Fluorescence can be monitored with an excitation wavelength of 544 nM and an emission wavelength of 572 nM. Reaction conditions will need to be optimized for each specific application. We recommend an initial Di-Ub (K48-linked) FRET TAMRA Pos1 concentration of 100-500 nM.
Source
E. coli-derived human Di-Ubiquitin protein
Accession #
Predicted Molecular Mass
17 kDa

Product Datasheets

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UF-210

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.

UF-210

Formulation X mg/ml (X μM) in 50 mM HEPES pH 7.5, 200 mM NaCl
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Protect from light. 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.
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Background: Di-Ubiquitin

With a predicted molecular weight of 17 kDa, Di-Ubiquitin is composed of two Ubiquitin monomers that are covalently linked through an isopeptide bond, which typically form between a lysine residue of one Ubiquitin molecule and the C-terminal glycine residue of another Ubiquitin molecule (1). Each human Ubiquitin monomer is 76 amino acids (aa) in length and shares 96% and 100% aa identity with yeast and mouse Ubiquitin, respectively (2). Ubiquitin has seven lysine residues that can participate in the formation of poly-Ubiquitin chains. The specific lysine residue used in Ubiquitin conjugation is thought to determine the function of poly-ubiquitination in cellular processes such as protein degradation, signaling, and trafficking (3-8).

Linkage specific di-Ubiquitin is a substrate for enzymes that cleave the isopeptide linkage between two Ubiquitin molecules. These FRET-based DUB substrates with isopeptide bonds between Ubiquitin moieties are superior to gel densitometry assays. Since DUBs recognize and cleave substrates with specific steric conditions, each substrate varies in the position of the fluorophore and quencher and must be empirically tested for individual enzymes.

References
  1. Scheffner, M. et al. (1995) Nature 373:81.
  2. Sharp, P.M. & W.-H. Li (1987) Trends Ecol. Evol. 2:328.
  3. Behrends, C. & J.W. Harper (2011) Nat. Struct. Mol. Biol. 18:520.
  4. Greene, W. et al. (2012) PLoS Pathog. 8:e1002703.
  5. Henry, A.G. et al. (2012) Dev. Cell 23:519.
  6. Tong, X. et al. (2012) J. Biol. Chem. 287:25280.
  7. Wei, W. et al. (2004) Nature 428:194.
  8. Zhang, J. et al. (2012) J. Biol. Chem. 287:28646.
Entrez Gene IDs
7314 (Human)
Alternate Names
DiUbiquitin; Di-Ubiquitin; HEL-S-50; Ub2; UBB; ubiquitin B

Citation for Recombinant Human Di-Ub (K48-linked) FRET TAMRA Pos1, 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.

1 Citation: Showing 1 - 1

  1. 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, 2021-05-19;0(0):.
    Species: Bacteria - Salmonella typhimurium
    Sample Types: Cell Lysates
    Applications: Bioassay

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