Recombinant Human Di-Ub/Ub2 WT Chains (K27-linked), CF

R&D Systems | Catalog # UC-61B

R&D Systems
Discontinued Product
UC-61B has been discontinued. View all Di-Ubiquitin products.

Key Product Details

Source

E. coli

Accession Number

P0CG47

Applications

Bioactivity
View all Di-Ubiquitin Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Di-Ubiquitin protein

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.  

 

Ub-AQUA analysis:

K27:   95.9%

K11:     3.9%

All other linkages ≤ 0.05%

Predicted Molecular Mass

17 kDa

Activity

Ubiquitin chains vary in length, linkage, and function. K27-linked Di-Ubiquitin Chains (Ub2) 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.

Formulation, Preparation, and Storage

UC-61B
Formulation 1 mg/ml (58 μM) in sterile, deionized water.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: Di-Ubiquitin

Linkage specific Poly-Ubiquitin chains may be used as a substrate for in vitro reactions with deubiquitinating enzymes ("DUB's") that cleave the peptide or isopeptide linkage between adjacent Ubiquitin molecules. Poly-Ubiquitin chains can also be used to investigate mechanisms of binding and recognition between the chains and other proteins that contain Ubiquitin-Associated domains (UBAs), Ubiquitin-interacting motifs (UIMs), ZnF's and/or other Ubiquitin-sensing elements.

K29-linked Di-Ubiquitin chains are manufactured using recombinant Ubiquitin and purely enzymatic techniques to avoid the potential for contaminating synthetic intermediates. The correctness of linkage and purity of each production lot is assessed using the Absolute Quantitation of Ubiquitin method (Ub-AQUA), an LCMS-based technique that provides extremely accurate information on the composition of Poly-Ubiquitin samples.

References

  1. Kirkpatrick D.S., et al. (2006) Nat Cell Biol. 8(7): 700-10
  2. Michel, M.A. et al. (2015) Mol Cell.  58(1): 95-109
  3. Ordureau, A., et al. (2014) Mol. Cell  56(3): 360–375
  4. Ordureau, A., et al. (2015) Pro. Nat. Acad. of Sci. USA 112(21): 6637–6642
  5. Phu L., et al. (2011) Mol Cell Proteomics 10(5): M110.003756

Alternate Names

DiUbiquitin, Ub2

Entrez Gene IDs

7314 (Human)

Gene Symbol

UBB

UniProt

P0CG47

Product Documents for Recombinant Human Di-Ub/Ub2 WT Chains (K27-linked), CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Di-Ub/Ub2 WT Chains (K27-linked), CF

For research use only

Related Research Areas

NFkB Pathway

Citations for Recombinant Human Di-Ub/Ub2 WT Chains (K27-linked), CF

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