AQUApure Tetra-Ub Chains (K63-linked) Protein, CF
AQUApure Tetra-Ub Chains (K63-linked) Protein, CF Summary
Product Specifications
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 Tetra-Ubiquitin Chains (Ub4) 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-310B
Formulation | 1 mg/ml (29 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: Tetra-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 Tetra-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: 700-10
- Ordureau, A. et al. (2014) Mol. Cell 56: 360–375
- Ordureau, A. et al. (2015) Pro. Nat. Acad. of Sci. USA 112: 6637–6642
- Phu L. et al. (2011) Mol Cell Proteomics 10: M110.003756
- Wall C.E. et al. (2019) Cell Reports 29: 3280
Citations for AQUApure Tetra-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.
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Citations: Showing 1 - 10
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Hepatitis B virus rigs the cellular metabolome to avoid innate immune recognition
Authors: L Zhou, R He, P Fang, M Li, H Yu, Q Wang, Y Yu, F Wang, Y Zhang, A Chen, N Peng, Y Lin, R Zhang, M Trilling, R Broering, M Lu, Y Zhu, S Liu
Nature Communications, 2021-01-04;12(1):98.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Sensitive ELISA-based detection method for the mitophagy marker p-S65-Ub in human cells, autopsy brain, and blood samples
Authors: JO Watzlawik, X Hou, D Fricova, C Ramnarine, SK Barodia, TF Gendron, MG Heckman, M DeTure, J Siuda, ZK Wszolek, CR Scherzer, OA Ross, G Bu, DW Dickson, MS Goldberg, FC Fiesel, W Springer
Autophagy, 2020-10-28;0(0):1-16.
Species: Human
Sample Types: N/A
Applications: Western Blot -
A feedforward circuit shaped by ECT2 and USP7 contributes to breast carcinogenesis
Authors: Q Zhang, C Cao, W Gong, K Bao, Q Wang, Y Wang, L Bi, S Ma, J Zhao, L Liu, S Tian, K Zhang, J Yang, Z Yao, N Song, L Shi
Theranostics, 2020-08-29;10(23):10769-10790.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
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 -
Regulation of HLTF-mediated PCNA polyubiquitination by RFC and PCNA monoubiquitination levels determines choice of damage tolerance pathway
Authors: Y Masuda, S Mitsuyuki, R Kanao, A Hishiki, H Hashimoto, C Masutani
Nucleic Acids Res., 2018-11-30;0(0):.
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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 -
Novel polyubiquitin imaging system, PolyUb-FC, reveals that K33-linked polyubiquitin is recruited by SQSTM1/p62
Authors: Y Nibe, S Oshima, M Kobayashi, C Maeyashiki, Y Matsuzawa, K Otsubo, H Matsuda, E Aonuma, Y Nemoto, T Nagaishi, R Okamoto, K Tsuchiya, T Nakamura, S Nakada, M Watanabe
Autophagy, 2018-01-24;0(0):1-43.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Non-canonical regulation of SPL transcription factors by a human OTUB1-like deubiquitinase defines a new plant type rice associated with higher grain yield
Authors: S Wang, K Wu, Q Qian, Q Liu, Q Li, Y Pan, Y Ye, X Liu, J Wang, J Zhang, S Li, Y Wu, X Fu
Cell Res., 2017-08-04;27(9):1142-1156.
Species: Plant - Oryza sativa (Rice)
Sample Types: Recombinant Protein
Applications: Bioassay -
Tandem UIMs confer Lys48 ubiquitin chain substrate preference to deubiquitinase USP25
Authors: K Kawaguchi, K Uo, T Tanaka, M Komada
Sci Rep, 2017-03-22;7(0):45037.
Applications: Bioassay
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