Recombinant Human Linear Tetra-Ubiquitin/Ub4 Protein, CF

Product Discontinued

Recombinant Human Linear Tetra-Ubiquitin/Ub4 Protein, CF Summary

• Details of Functionality: Ubiquitin chains vary in length, linkage, and function. Linear Tetra-Ubiquitin Chains 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: AQUApure 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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 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
• Reconstitution Instructions: Reconstitute at 2 mg/mL in an aqueous solution.

Notes

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

Alternate Names for Recombinant Human Linear Tetra-Ubiquitin/Ub4 Protein, 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).

This linear Ubiquitin fusion protein can be used as a substrate for deubiquitinating enzymes (DUBs) to test for non-isopeptide bond cleavage activity or preference. Ub is not expressed directly as free Ub but rather as linear fusions either to itself or to certain ribosomal protein subunits. These Ub-fusion precursors are proteolyzed by deubiquitinating enzymes (DUBs) at the appropriate junction points to yield active Ub monomers with C-termini ending in GG. There are likely several intracellular DUBs which perform this essential processing role. This protein can also be used to investigate the mechanism of binding and recognition by E1 or E2 enzymes, DUBs, E3 ligases or other proteins that contain Ubiquitin-associated domains (UBAs) or Ubiquitin-interacting motifs (UIMs).

 

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 (UC-710)(3)

Publications using UC-710

• JO Watzlawik, X Hou, D Fricova, C Ramnarine, SK Barodia, TF Gendron, MG Heckman, M DeTure, J Siuda, ZK Wszolek, CR Scherzer, OA Ross, G Bu, DW Dickson, MS Goldberg, FC Fiesel, W Springer Sensitive ELISA-based detection method for the mitophagy marker p-S65-Ub in human cells, autopsy brain, and blood samples Autophagy, 2020-10-28;0(0):1-16. 2020-10-28 [PMID: 33112198] (Western Blot, Human)
• P Haahr, N Borgermann, X Guo, D Typas, D Achuthanku, S Hoffmann, R Shearer, TK Sixma, N Mailand ZUFSP Deubiquitylates K63-Linked Polyubiquitin Chains to Promote Genome Stability Mol. Cell, 2018-03-22;0(0):. 2018-03-22 [PMID: 29576528] (Bioassay, Human)
• H Takeda, W Zhou, K Kido, R Suno, T Iwasaki, T Kobayashi, T Sawasaki CP5 system, for simple and highly efficient protein purification with a C-terminal designed mini tag PLoS ONE, 2017-05-25;12(5):e0178246. 2017-05-25 [PMID: 28542437] (Enzyme Assay)

Bioinformatics

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