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

• Reactivity: All-Multi
• Applications: Bioactivity
• Format: Carrier-Free

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

• Additional Information: Will be discontinued when existing inventory is gone.
• 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 degreesC for 20 minutes prior to gel electrophoresis.
• Source: E. coli-derived Di-Ubiquitin protein
• Accession #: P0CG47
• 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

• Dilutions:
  • Bioactivity
• 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 degreesC as supplied. 3 months, -20 to -70 degreesC 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

Publications for Di-Ubiquitin (UC-11B)(9)

Publications using UC-11B

• Y Shi, H Liu, K Ma, ZQ Luo, J Qiu Legionella longbeachae effector protein RavZ inhibits autophagy and regulates phagosome ubiquitination during infection PLoS ONE, 2023-02-09;18(2):e0281587. 2023-02-09 [PMID: 36758031] (Bioassay, Legionella)
• N Manczyk, G Veggiani, J Teyra, AW Strilchuk, SS Sidhu, F Sicheri The ubiquitin interacting motifs of USP37 act on the proximal Ub of a di-Ub chain to enhance catalytic efficiency Sci Rep, 2019-03-11;9(1):4119. 2019-03-11 [PMID: 30858488] (Bioassay)
• P Haahr, N Borgermann, X Guo, D Typas, D Achuthanku, S Hoffmann, R Shearer, TK Sixma, N Mailand ZUFSP Deubiquitylates K63-Linked Polyubiquitin Chains to Promote Genome Stability Mol. Cell, 2018-03-22;0(0):. 2018-03-22 [PMID: 29576528] (Bioassay, Human)
• S Rasool, N Soya, L Truong, N Croteau, GL Lukacs, JF Trempe PINK1 autophosphorylation is required for ubiquitin recognition EMBO Rep., 2018-02-23;0(0):. 2018-02-23 [PMID: 29475881] (Bioassay, Insect - Tribolium castaneum)
• Y Zhao, MC Mudge, JM Soll, RB Rodrigues, AK Byrum, EA Schwarzkop, TR Bradstreet, SP Gygi, BT Edelson, N Mosammapar OTUD4 Is a Phospho-Activated K63 Deubiquitinase that Regulates MyD88-Dependent Signaling Mol. Cell, 2018-02-01;69(3):505-516.e5. 2018-02-01 [PMID: 29395066] (Bioassay, Human)
• H Zheng, M Wang, C Zhao, S Wu, P Yu, Y Lü, T Wang, Y Ai Characterization of the deubiquitination activity and substrate specificity of the chicken ubiquitin-specific protease 1/USP associated factor 1 complex PLoS ONE, 2017-11-01;12(11):e0186535. 2017-11-01 [PMID: 29091922] (Bioassay, Human)
• 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 Cholesterol and fatty acids regulate cysteine ubiquitylation of ACAT2 through competitive oxidation Nat. Cell Biol., 2017-06-12;19(7):808-819. 2017-06-12 [PMID: 28604676] (Control)
• H Tsuchiya, F Ohtake, N Arai, A Kaiho, S Yasuda, K Tanaka, Y Saeki In�Vivo Ubiquitin Linkage-type Analysis Reveals that the Cdc48-Rad23/Dsk2 Axis Contributes to K48-Linked Chain Specificity of the Proteasome Mol. Cell, 2017-05-18;66(4):488-502.e7. 2017-05-18 [PMID: 28525741] (Bioassay)
• F He, HP Wollscheid, U Nowicka, M Biancospin, E Valentini, A Ehlinger, F Acconcia, E Magistrati, S Polo, KJ Walters Myosin VI Contains a Compact Structural Motif that Binds to Ubiquitin Chains Cell Rep, 2016-03-10;14(11):2683-94. 2016-03-10 [PMID: 26971995] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • P0CG47()