Recombinant Human Di-Ub (K63-linked) FRET TAMRA Pos3, CF Summary
Product Datasheets
Carrier Free
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.
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-330
| Formulation | X mg/ml (X µM) in 50 mM HEPES pH 7.5, 200 mM NaCl, 2 mM DTT |
| 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.
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Reconstitution Calculator
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 andmouse 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.
- Scheffner, M. et al. (1995) Nature 373:81.
- Sharp, P.M. & W.-H. Li (1987) Trends Ecol. Evol. 2:328.
- Behrends, C. & J.W. Harper (2011) Nat. Struct. Mol. Biol. 18:520.
- Greene, W. et al. (2012) PLoS Pathog. 8:e1002703.
- Henry, A.G. et al. (2012) Dev. Cell 23:519.
- Tong, X. et al. (2012) J. Biol. Chem. 287:25280.
- Wei, W. et al. (2004) Nature 428:194.
- Zhang, J. et al. (2012) J. Biol. Chem. 287:28646.
Citations for Recombinant Human Di-Ub (K63-linked) FRET TAMRA Pos3, 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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- The AMSH3 ESCRT-III-Associated Deubiquitinase Is Essential for Plant ImmunityAuthors: T Schultz-La, A Lenk, K Kalinowska, LK Vestergaar, C Pedersen, E Isono, H Thordal-ChCell Rep, 2018;25(9):2329-2338.e5.Species: Plant - Arabidopsis thalianaSample Types: Recombinant ProteinApplications: Ubiquitination
- Arabidopsis SH3P2 is an ubiquitin-binding protein that functions together with ESCRT-I and the deubiquitylating enzyme AMSH3Authors: MK Nagel, K Kalinowska, K Vogel, GD Reynolds, Z Wu, F Anzenberge, M Ichikawa, C Tsutsumi, MH Sato, B Kuster, SY Bednarek, E IsonoProc. Natl. Acad. Sci. U.S.A., 2017;0(0):.Species: N/ASample Types: Recombinant ProteinApplications: Bioassay
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