Recombinant Human Di-Ub/Ub2 WT Chains (K27-linked), CF
Recombinant Human Di-Ub/Ub2 WT Chains (K27-linked), CF Summary
Product Specifications
Ub-AQUA analysis:
K27: 95.9%
K11: 3.9%
All other linkages ≤ 0.05%
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.
Product Datasheets
Carrier Free
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.
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.
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.
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Reconstitution Calculator
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.
- Kirkpatrick D.S., et al. (2006) Nat Cell Biol. 8(7): 700-10
- Michel, M.A. et al. (2015) Mol Cell. 58(1): 95-109
- Ordureau, A., et al. (2014) Mol. Cell 56(3): 360–375
- Ordureau, A., et al. (2015) Pro. Nat. Acad. of Sci. USA 112(21): 6637–6642
- Phu L., et al. (2011) Mol Cell Proteomics 10(5): M110.003756
Citations for Recombinant Human Di-Ub/Ub2 WT Chains (K27-linked), 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.
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Citations: Showing 1 - 10
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The ubiquitin interacting motifs of USP37 act on the proximal Ub of a di-Ub chain to enhance catalytic efficiency
Authors: N Manczyk, G Veggiani, J Teyra, AW Strilchuk, SS Sidhu, F Sicheri
Sci Rep, 2019-03-11;9(1):4119.
Applications: Bioassay -
USP52 acts as a deubiquitinase and promotes histone chaperone ASF1A stabilization
Authors: S Yang, L Liu, C Cao, N Song, Y Wang, S Ma, Q Zhang, N Yu, X Ding, F Yang, S Tian, K Zhang, T Sun, J Yang, Z Yao, S Wu, L Shi
Nat Commun, 2018-03-29;9(1):1285.
Applications: Bioassay -
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, 2018-03-22;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Bioassay -
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, 2018-02-01;69(3):505-516.e5.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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, 2017-11-01;12(11):e0186535.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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, 2017-05-18;66(4):488-502.e7.
Applications: Bioassay -
ER protein SCAP inhibits Dengue virus NS2B3 protease by suppressing its K27-linked polyubiquitylation
Authors: H Liu, L Zhang, J Sun, W Chen, S Li, Q Wang, H Yu, Z Xia, X Jin, C Wang
J. Virol, 2017-04-13;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
A PALB2-interacting domain in RNF168 couples homologous recombination to DNA break-induced chromatin ubiquitylation
Authors: MS Luijsterbu, D Typas, MC Caron, WW Wiegant, D van den He, RA Boonen, AM Couturier, LH Mullenders, JY Masson, H van Attiku
Elife, 2017-02-27;6(0):.
Applications: Bioassay -
Myosin VI Contains a Compact Structural Motif that Binds to Ubiquitin Chains
Authors: F He, HP Wollscheid, U Nowicka, M Biancospin, E Valentini, A Ehlinger, F Acconcia, E Magistrati, S Polo, KJ Walters
Cell Rep, 2016-03-10;14(11):2683-94.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Deubiquitinase-based analysis of ubiquitin chain architecture using Ubiquitin Chain Restriction (UbiCRest).
Authors: Hospenthal, Manuela, Mevissen, Tycho E, Komander, David
Nat Protoc, 2015-01-29;10(2):349-61.
Species: Human
Sample Types: Protein
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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, 2014-12-23;34(3):326-43.
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Specific recognition of linear polyubiquitin by A20 zinc finger 7 is involved in NF-kappaB regulation.
EMBO J., 2012-08-28;31(19):3856-70.
Species: Human
Sample Types: Recombinant Protein
Applications: Binding Assay
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