AQUApure Di-Ub Chains (K6-linked) Protein, CF

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Product Details

Summary
Reactivity MultiSpecies Glossary
Applications Bioactivity
Format
Carrier-Free

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AQUApure Di-Ub Chains (K6-linked) Protein, CF Summary

Details of Functionality

Ubiquitin chains vary in length, linkage, and function. K6-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.

Source
E. coli-derived Di-Ubiquitin protein
Accession #
Protein/Peptide Type
AQUApure Recombinant Proteins
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

 

Ub-AQUA analysis:

K6  :   99.19%

K63:     0.40%

K48:     0.22%

K11:     0.14%

All other linkages ≤ 0.03%

 

Applications/Dilutions

Theoretical MW
17 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 opening.
Buffer
1 mg/ml (58 μM) in sterile, deionized water
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain.

 

Ub-AQUA analysis:

K6  :   99.19%

K63:     0.40%

K48:     0.22%

K11:     0.14%

All other linkages ≤ 0.03%

 

Notes

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

Alternate Names for AQUApure Di-Ub Chains (K6-linked) Protein, CF

  • DiUbiquitin
  • Di-Ubiquitin
  • HEL-S-50
  • Ub2
  • UBB
  • ubiquitin B

Background

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.

K6-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.

 

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

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UC-11B
Species: Multi
Applications: Bioactivity

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