Recombinant Human His6-PolyUb WT Chains (2-7,K63-linked), CF

Product Discontinued

Recombinant Human His6-PolyUb WT Chains (2-7,K63-linked), CF Summary

• Details of Functionality: Ubiquitin chains vary in length, linkage, and function. K63-linked His6-Poly-Ubiquitin Chains (Ub2-7) 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 Poly-Ubiquitin protein
  Contains a 6-His tag
• Accession #: P0CG47
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 19 kDa (Ub2), 29 kDa (Ub3), 38 kDa (Ub4), 48 kDa (Ub5), 57 kDa (Ub6), and 67 kDa (Ub7).
  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 opening.
• Buffer: X mg/ml in sterile, 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 His6-PolyUb WT Chains (2-7,K63-linked), CF

• CEP80
• HEL112
• PolyUbiquitin
• Poly-Ubiquitin
• ribosomal protein S27a
• RPS27A
• S27A
• UBA80
• UBC
• UBCEP1
• UBCEP80

Background

Poly-Ubiquitin chains are composed of 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).

Linkage specific poly-Ubiquitin chains are used to investigate mechanisms of chain recognition, binding and hydrolysis by the proteasome, deubiquitinating enzymes, E3 ligases or other proteins that contain Ubiquitin-associated domains (UBAs) or ubiquitin-interacting motifs (UIMs). Lys63-linked poly-Ubiquitin has been implicated in several non-degradative processes such as receptor endocytosis and sorting, translation, DNA damage repair, the stress response and signaling in theNF kappa B pathway. This product is formed with wild-type human recombinant Ubiquitin and linkage-specific enzymes. This mixture of poly-Ubiquitin chains contains di-Ubiquitin and higher MW species; mono-Ubiquitin has been removed.

The His6-tag is convenient for metal chelate affinity purification and immuno-detection using His6-specific antibodies.

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 Poly-Ubiquitin (UCH-330)(1)

Publication using UCH-330

• C Sheng, C Yao, Z Wang, H Chen, Y Zhao, D Xu, H Huang, W Huang, S Chen Cyclophilin J limits inflammation through the blockage of ubiquitin chain sensing Nat Commun, 2018-10-22;9(1):4381. 2018-10-22 [PMID: 30348973] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • P0CG47()