AQUApure Di-Ub Chains (K63-linked) Protein, CF

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Citations (12)
Reviews (2)

AQUApure Di-Ub Chains (K63-linked) Protein, CF Summary

Product Specifications

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


Ub-AQUA analysis:

K63:   99.04%

K11:     0.76%

K6  :     0.13%

All other linkages ≤ 0.04%


Ubiquitin chains vary in length, linkage, and function. K63-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.

E. coli-derived human Di-Ubiquitin protein
Accession #
Predicted Molecular Mass
17 kDa

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.


Formulation 1 mg/ml (58 μM) in sterile, deionized water
Shipping The product is shipped with dry ice or equivalent. 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.
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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.

K63-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
Entrez Gene IDs
7314 (Human)
Alternate Names
DiUbiquitin; Di-Ubiquitin; HEL-S-50; Ub2; UBB; ubiquitin B

Citations for AQUApure Di-Ub Chains (K63-linked) Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

12 Citations: Showing 1 - 10
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  1. Structural insights into the mechanism and inhibition of transglutaminase-induced ubiquitination by the Legionella effector MavC
    Authors: Y Mu, Y Wang, Y Huang, D Li, Y Han, M Chang, J Fu, Y Xie, J Ren, H Wang, Y Zhang, ZQ Luo, Y Feng
    Nat Commun, 2020;11(1):1774.
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Bioassay
  2. Post-translational Modification of OTULIN Regulates Ubiquitin Dynamics and Cell Death
    Authors: T Douglas, M Saleh
    Cell Rep, 2019;29(11):3652-3663.e5.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  3. Reactive-site-centric chemoproteomics identifies a distinct class of deubiquitinase enzymes
    Authors: DS Hewings, J Heideker, TP Ma, AP AhYoung, F El Oualid, A Amore, GT Costakes, D Kirchhofer, B Brasher, T Pillow, N Popovych, T Maurer, C Schwerdtfe, WF Forrest, K Yu, J Flygare, M Bogyo, IE Wertz
    Nat Commun, 2018;9(1):1162.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  4. ZUFSP Deubiquitylates K63-Linked Polyubiquitin Chains to Promote Genome Stability
    Authors: P Haahr, N Borgermann, X Guo, D Typas, D Achuthanku, S Hoffmann, R Shearer, TK Sixma, N Mailand
    Mol. Cell, 2018;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Bioassay
  5. OTUD4 Is a Phospho-Activated K63 Deubiquitinase that Regulates MyD88-Dependent Signaling
    Authors: Y Zhao, MC Mudge, JM Soll, RB Rodrigues, AK Byrum, EA Schwarzkop, TR Bradstreet, SP Gygi, BT Edelson, N Mosammapar
    Mol. Cell, 2018;69(3):505-516.e5.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  6. PINK1 autophosphorylation is required for ubiquitin recognition
    Authors: S Rasool, N Soya, L Truong, N Croteau, GL Lukacs, JF Trempe
    EMBO Rep., 2018;0(0):.
    Species: Insect - Tribolium castaneum
    Sample Types: Recombinant Protein
    Applications: Bioassay
  7. Cholesterol and fatty acids regulate cysteine ubiquitylation of ACAT2 through competitive oxidation
    Authors: YJ Wang, Y Bian, J Luo, M Lu, Y Xiong, SY Guo, HY Yin, X Lin, Q Li, CCY Chang, TY Chang, BL Li, BL Song
    Nat. Cell Biol., 2017;19(7):808-819.
    Species: N/A
    Sample Types:
    Applications: Control
  8. In�Vivo Ubiquitin Linkage-type Analysis Reveals that the Cdc48-Rad23/Dsk2 Axis Contributes to K48-Linked Chain Specificity of the Proteasome
    Authors: H Tsuchiya, F Ohtake, N Arai, A Kaiho, S Yasuda, K Tanaka, Y Saeki
    Mol. Cell, 2017;66(4):488-502.e7.
    Applications: Bioassay
  9. Characterization of the deubiquitination activity and substrate specificity of the chicken ubiquitin-specific protease 1/USP associated factor 1 complex
    Authors: H Zheng, M Wang, C Zhao, S Wu, P Yu, Y Lü, T Wang, Y Ai
    PLoS ONE, 2017;12(11):e0186535.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  10. Deubiquitinase-based analysis of ubiquitin chain architecture using Ubiquitin Chain Restriction (UbiCRest).
    Authors: Hospenthal, Manuela, Mevissen, Tycho E, Komander, David
    Nat Protoc, 2015;10(2):349-61.
    Species: Human
    Sample Types: Protein
  11. USP45 deubiquitylase controls ERCC1-XPF endonuclease-mediated DNA damage responses.
    Authors: Perez-Oliva A, Lachaud C, Szyniarowski P, Munoz I, Macartney T, Hickson I, Rouse J, Alessi D
    EMBO J, 2015;34(3):326-43.
  12. The vOTU domain of highly-pathogenic porcine reproductive and respiratory syndrome virus displays a differential substrate preference.
    Authors: Deaton M, Spear A, Faaberg K, Pegan S
    Virology, 2014;454(0):247-53.


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Reviews for AQUApure Di-Ub Chains (K63-linked) Protein, CF

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AQUApure Di-Ub Chains (K63-linked) Protein, CF
By Anonymous on 12/17/2019
Application: Biochemistry Course
Reason for Rating: The faculty were pleased with the outcome.

AQUApure Di-Ub Chains (K63-linked) Protein, CF
By Anonymous on 07/02/2019
Application: Enzymatic activity in vitro