Recombinant Human Tetra-Ub/Ub4 WT Chains (K48/K63/K48), CF

Product Discontinued

Recombinant Human Tetra-Ub/Ub4 WT Chains (K48/K63/K48), CF Summary

• Details of Functionality: Ubiquitin chains vary in length, linkage, and function.Tetra-Ubiquitin Chains (Ub-K48-Ub-K63-Ub-K48-Ub) are ideal for investigating Ubiquitin-binding proteins and as substrates for Ubiquitin-specific isopeptidases. 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 Tetra-Ubiquitin protein
• Accession #: P0CG47
• Protein/Peptide Type: Recombinant Proteins
• Gene: UBB
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 34 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 24 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a solution in deionized water.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Tetra-Ub/Ub4 WT Chains (K48/K63/K48), CF

• HEL-S-50
• TetraUbiquitin
• Tetra-Ubiquitin
• Ub4
• UBB
• ubiquitin B

Background

With a predicted molecular weight of 34 kDa, Tetra-Ubiquitin chains are composed of four Ubiquitin monomers that are covalently linked through  isopeptide bonds, 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). Seven of the 76 aa in Ubiquitin are lysine residues that can participate in poly-Ubiquitin chain formation. Linkage through specific lysine residues is thought to serve as a signal that affects protein degradation, signaling, trafficking, and other cellular processes (3-8).

Tetra-Ub can be used to investigate mechanisms of binding and recognition by deubiquitinating enzymes, E3 ligases or other proteins that contain Ubiquitin-associated domains (UBAs) or Ubiquitin-interacting motifs (UIMs). Tetra-Ub is the minimal unit necessary for recognition by the 26S Proteasome and contains structural characteristic (such as repeating hydrophobic patches) not present in di-Ub. This product is made with wild-type human recombinant Ubiquitin and linkage-specific enzymes, which results in one K63-linkage between two K48-linkages.

 

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.

Publications for Tetra-Ubiquitin (UCM-310)(1)

Publication using UCM-310

• M Deng, J Lin, S Nowsheen, T Liu, Y Zhao, PW Villalta, D Sicard, DJ Tschumperl, S Lee, J Kim, Z Lou Extracellular matrix stiffness determines DNA repair efficiency and cellular sensitivity to genotoxic agents Science Advances, 2020-09-11;6(37):. 2020-09-11 [PMID: 32917705] (Bioassay, Human)

Bioinformatics

• Gene Symbol: UBB
• Uniprot:
  • P0CG47()