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Recombinant Human Di-Ub/Ub2 Non-hydrolyzable (K48-linked) CF

Catalog #: UCN-200 Datasheet / COA / SDS

Discontinued Product

UCN-200 has been discontinued.
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Recombinant Human Di-Ub/Ub2 Non-hydrolyzable (K48-linked) CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain
Activity
Ubiquitin chains vary in length, linkage, and function. K48-linked Non-hydrolyzable Di-Ubiquitin Chains (Ub2) may be useful for investigating Ubiquitin-binding proteins and exploring the role of unanchored Ubiquitin chains in signaling pathways. Reaction conditions will need to be optimized for each specific application. IMPORTANT: Heating this product in SDS-PAGE buffer or terminating reactions containing this product with heated SDS-PAGE buffer could lead to unexpected, high apparent molecular weight banding or smearing on gels that is not representative of product purity. For optimal results, we recommend incubation in SDS-PAGE buffer + DTT at <40 °C for 20 minutes prior to gel electrophoresis.
Source
E. coli-derived human Di-Ubiquitin protein

Each Ubiquitin contains a Pro substitution at position 73.

Accession #
P0CG47
Predicted Molecular Mass
17 kDa

Product Datasheets

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UCN-200

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.

UCN-200

Formulation Lyophilized from a solution in deionized water.
Reconstitution Reconstitute at 5 mg/ml in aqueous buffer.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 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: 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, non-hydrolyzable di-Ubiquitin is resistant to the activity of deubiquitinating enzymes (DUB's) that cleave the isopeptide linkage between adjacent Ubiquitin molecules. It can be used to investigate binding interactions between di-Ubiquitin and proteins that contain elements such as Ubiquitin-associated domains (UBAs) or Ubiquitin-interacting motifs (UIMs). This product may also be useful in exploring the role of unanchored poly-Ubiquitin chains in some signaling pathways.

 

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

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