Recombinant Human Ubiquitin AMC Protein, CF

Catalog # Availability Size / Price Qty
U-550-050
Product Details
Citations (35)
FAQs
Reviews (1)

Recombinant Human Ubiquitin AMC Protein, CF Summary

Purity
>95%, by HPLC.
Activity
Recombinant Human Ubiquitin-AMC is a fluorogenic substrate for some Ubiquitin-specific isopeptidases. Release of AMC fluorescence can be monitored with an excitation wavelength of 345 nm and an emission wavelength of 445 nm. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human Ubiquitin-AMC concentration of 0.1-1 μM.
Source
E. coli-derived human Ubiquitin protein
Accession #
Predicted Molecular Mass
8.7 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.

U-550

Formulation

2.2 mg/ml (250 µM) in 100% DMSO

Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
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Background: Ubiquitin

Ubiquitin is a 76 amino acid (aa) protein that is ubiquitously expressed in all eukaryotic organisms. Ubiquitin is highly conserved with 96% aa sequence identity shared between human and yeast Ubiquitin, and 100% aa sequence identity shared between human and mouse Ubiquitin (1). In mammals, four Ubiquitin genes encode for two Ubiquitin-ribosomal fusion proteins and two poly-Ubiquitin proteins. Cleavage of the Ubiquitin precursors by deubiquitinating enzymes gives rise to identical Ubiquitin monomers each with a predicted molecular weight of 8.6 kDa. Conjugation of Ubiquitin to target proteins involves the formation of an isopeptide bond between the C-terminal glycine residue of Ubiquitin and a lysine residue in the target protein. This process of conjugation, referred to as ubiquitination or ubiquitylation, is a multi-step process that requires three enzymes: a Ubiquitin-activating (E1) enzyme, a Ubiquitin-conjugating (E2) enzyme, and a Ubiquitin ligase (E3). Ubiquitination is classically recognized as a mechanism to target proteins for degradation and as a result, Ubiquitin was originally named ATP-dependent Proteolysis Factor 1 (APF-1) (2,3). In addition to protein degradation, ubiquitination has been shown to mediate a variety of biological processes such as signal transduction, endocytosis, and post-endocytic sorting (4-7).

This fluorogenic substrate for Ubiquitin hydrolases is based on the C-terminus derivatization of Ubiquitin with 7-amino-4-methylcoumarin (AMC). Ubiquitin-AMC is an exquisitely sensitive substrate for UCHL3 (Km = 0.039 μM) and for Isopeptidase-T (Km = 0.17 - 1.4 μM). Ubiquitin-AMC is useful for studying Ubiquitin hydrolases when detection sensitivity or continuous monitoring of activity is essential.

References
  1. Sharp, P.M. & W.-H. Li. (1987) Trends Ecol. Evol. 2:328.
  2. Ciechanover, A. et al. (1980 ) Proc. Natl. Acad. Sci. USA 77:1365.
  3. Hershko, A. et al. (1980) Proc. Natl. Acad. Sci. USA 77:1783.
  4. Greene, W. et al. (2012) PLoS Pathog. 8:e1002703.
  5. Tong, X. et al. (2012) J. Biol. Chem. 287:25280.
  6. Wei, W. et al. (2004) Nature 428:194.
  7. Wertz, I.E. et al. (2004) Nature 430:694.
  8. Dang L.C., et al. (1998) Biochem. 37:1868-1879.
  9. Mason D.E., et al. (2004) Biochem 43:6535-6544.
  10. Stein R.L., et al. (1998) Biochem. 34:12616-12623.  
Entrez Gene IDs
7314 (Human); 298693 (Rat)
Alternate Names
RPS27A; UBA52; UBB ubiquitin B; UBB; UBC; Ubiquitin

Citations for Recombinant Human Ubiquitin AMC 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.

35 Citations: Showing 1 - 10
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  1. Nedd8 hydrolysis by UCH proteases in Plasmodium parasites
    Authors: M Karpiyevic, S Adjalley, M Mol, DB Ascher, B Mason, GJ van der He, H Laman, H Ovaa, MCS Lee, K Artavanis-
    PLoS Pathog., 2019;15(10):e1008086.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Characterizing the PRRSV nsp2 Deubiquitinase Reveals Dispensability of Cis-Activity for Replication and a Link of nsp2 to Inflammation Induction
    Authors: S Zhou, X Ge, C Kong, T Liu, A Liu, P Gao, J Song, L Zhou, X Guo, J Han, H Yang
    Viruses, 2019;11(10):.
    Species: Virus - PRRSV
    Sample Types: Recombinant Protein
    Applications: Bioassay
  3. Stable Occupancy of the Crimean-Congo Hemorrhagic Fever Virus-Encoded Deubiquitinase Blocks Viral Infection
    Authors: FEM Scholte, BL Hua, JR Spengler, JV Dzimianski, JD Coleman-Mc, SR Welch, LK McMullan, ST Nichol, SD Pegan, CF Spiropoulo, É Bergeron
    MBio, 2019;10(4):.
    Species: Virus
    Sample Types: Protein
    Applications: Bioassay
  4. Ubiquitination of UVRAG by SMURF1 promotes autophagosome maturation and inhibits hepatocellular carcinoma growth
    Authors: X Feng, Y Jia, Y Zhang, F Ma, Y Zhu, X Hong, Q Zhou, R He, H Zhang, J Jin, D Piao, H Huang, Q Li, X Qiu, Z Zhang
    Autophagy, 2019;0(0):1-20.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  5. Crystal structure and activity-based labeling reveal the mechanisms for linkage-specific substrate recognition by deubiquitinase USP9X
    Authors: P Paudel, Q Zhang, C Leung, HC Greenberg, Y Guo, YH Chern, A Dong, Y Li, M Vedadi, Z Zhuang, Y Tong
    Proc. Natl. Acad. Sci. U.S.A., 2019;0(0):.
    Applications: Bioassay
  6. 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;9(1):4119.
    Applications: Bioassay
  7. Ubiquitin C-Terminal Hydrolase L1 regulates autophagy by inhibiting autophagosome formation through its deubiquitinating enzyme activity
    Authors: C Yan, H Huo, C Yang, T Zhang, Y Chu, Y Liu
    Biochem. Biophys. Res. Commun., 2018;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  8. Ubiquitin C-Terminal Hydrolase L1 regulates autophagy by inhibiting autophagosome formation through its deubiquitinating enzyme activity
    Authors: C Yan, H Huo, C Yang, T Zhang, Y Chu, Y Liu
    Biochem. Biophys. Res. Commun., 2018;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  9. 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
  10. A new gold(I) complex-Au(PPh3)PT is a deubiquitinase inhibitor and inhibits tumor growth
    Authors: X Li, Q Huang, H Long, P Zhang, H Su, J Liu
    EBioMedicine, 2018;0(0):.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  11. USP15-dependent lysosomal pathway controls p53-R175H turnover in ovarian cancer cells
    Authors: A Padmanabha, N Candelaria, KK Wong, BC Nikolai, DM Lonard, BW O'Malley, JS Richards
    Nat Commun, 2018;9(1):1270.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  12. The novel anti-androgen candidate galeterone targets deubiquitinating enzymes, USP12 and USP46, to control prostate cancer growth and survival
    Authors: UL McClurg, M Azizyan, DT Dransfield, N Namdev, NCTH Chit, S Nakjang, CN Robson
    Oncotarget, 2018;9(38):24992-25007.
    Applications: Bioassay
  13. TRIM11 activates the proteasome and promotes overall protein degradation by regulating USP14
    Authors: L Chen, G Zhu, EM Johns, X Yang
    Nat Commun, 2018;9(1):1223.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  14. Bilirubin neurotoxicity is associated with proteasome inhibition
    Authors: H Huang, M Guo, N Liu, C Zhao, H Chen, X Wang, S Liao, P Zhou, Y Liao, X Chen, X Lan, J Chen, D Xu, X Li, X Shi, L Yu, Y Nie, X Wang, CE Zhang, J Liu
    Cell Death Dis, 2017;8(6):e2877.
    Species: N/A
    Sample Types: Protein
    Applications: Enzyme Assay
  15. 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
  16. Generation and Validation of Intracellular Ubiquitin Variant Inhibitors for USP7 and USP10
    Authors: W Zhang, MA Sartori, T Makhnevych, KE Federowicz, X Dong, L Liu, S Nim, A Dong, J Yang, Y Li, D Haddad, A Ernst, D Heerding, Y Tong, J Moffat, SS Sidhu
    J. Mol. Biol., 2017;0(0):.
    Applications: Bioassay
  17. An inhibitor of the proteasomal deubiquitinating enzyme USP14 induces tau elimination in cultured neurons
    Authors: M Boselli, BH Lee, J Robert, MA Prado, SW Min, C Cheng, MC Silva, C Seong, S Elsasser, KM Hatle, TC Gahman, SP Gygi, SJ Haggarty, L Gan, RW King, D Finley
    J. Biol. Chem., 2017;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  18. A mobile loop near the active site acts as a switch between the dual activities of a viral protease/deubiquitinase
    Authors: I Jupin, M Ayach, L Jomat, S Fieulaine, S Bressanell
    PLoS Pathog., 2017;13(11):e1006714.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  19. A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibility
    Authors: JF Beckmann, JA Ronau, M Hochstrass
    Nat Microbiol, 2017;2(0):17007.
    Species: N/A
    Sample Types: Protein
    Applications: Bioassay
  20. Ubiquitin Recognition of BAP1: Understanding its Enzymatic Function
    Authors: P Hanpude, S Bhattachar, AK Singh, TK Maiti
    Biosci. Rep., 2017;0(0):.
    Species: N/A
    Sample Types: Protein
    Applications: Enzyme Assay
  21. Chlamydia trachomatis-containing vacuole serves as deubiquitination platform to stabilize Mcl-1 and to interfere with host defense
    Authors: A Fischer, KS Harrison, Y Ramirez, D Auer, SR Chowdhury, BK Prusty, F Sauer, Z Dimond, C Kisker, P Scott Heft, T Rudel
    Elife, 2017;6(0):.
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  22. Ubiquitin Recognition of BAP1: Understanding its Enzymatic Function
    Authors: P Hanpude, S Bhattachar, AK Singh, TK Maiti
    Biosci. Rep., 2017;0(0):.
    Species: N/A
    Sample Types: Protein
    Applications: Enzyme Assay
  23. Diverse mechanisms of metaeffector activity in an intracellular bacterial pathogen, Legionella pneumophila
    Authors: Malene L Urbanus
    Mol. Syst. Biol, 2016;12(12):893.
    Applications: Bioassay
  24. Saturation scanning of ubiquitin variants reveals a common hot spot for binding to USP2 and USP21
    Proc Natl Acad Sci USA, 2016;113(31):8705-10.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay
  25. The proteasome deubiquitinase inhibitor VLX1570 shows selectivity for ubiquitin-specific protease-14 and induces apoptosis of multiple myeloma cells
    Sci Rep, 2016;6(0):26979.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  26. p53 down-regulates SARS coronavirus replication and is targeted by the SARS-unique domain and PLpro via E3 ubiquitin ligase RCHY1
    Proc Natl Acad Sci USA, 2016;113(35):E5192-201.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  27. BBX21, an Arabidopsis B-box protein, directly activates HY5 and is targeted by COP1 for 26S proteasome-mediated degradation
    Authors: Dongqing Xu
    Proc Natl Acad Sci USA, 2016;0(0):.
    Species: N/A
    Sample Types: Protein
    Applications: Bioassay
  28. Deubiquitination and Activation of AMPK by USP10
    Authors: M Deng, X Yang, B Qin, T Liu, H Zhang, W Guo, SB Lee, JJ Kim, J Yuan, H Pei, L Wang, Z Lou
    Mol. Cell, 2016;61(4):614-24.
    Species: N/A
    Sample Types: Protein
    Applications: Enzyme Assay
  29. X-ray Structural and Functional Studies of the Three Tandemly Linked Domains of Non-structural Protein 3 (nsp3) from Murine Hepatitis Virus Reveal Conserved Functions.
    Authors: Chen Y, Savinov S, Mielech A, Cao T, Baker S, Mesecar A
    J Biol Chem, 2015;290(42):25293-306.
    Species: N/A
    Sample Types: Protein
    Applications: Enzyme Assay
  30. The human otubain2-ubiquitin structure provides insights into the cleavage specificity of poly-ubiquitin-linkages.
    Authors: Altun M, Walter T, Kramer H, Herr P, Iphofer A, Bostrom J, David Y, Komsany A, Ternette N, Navon A, Stuart D, Ren J, Kessler B
    PLoS ONE, 2015;10(1):e0115344.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  31. Small-molecule RA-9 inhibits proteasome-associated DUBs and ovarian cancer in vitro and in vivo via exacerbating unfolded protein responses.
    Authors: Coughlin K, Anchoori R, Iizuka Y, Meints J, MacNeill L, Vogel R, Orlowski R, Lee M, Roden R, Bazzaro M
    Clin Cancer Res, 2014;20(12):3174-86.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay
  32. 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.
  33. Dopamine D3 receptor inhibits the ubiquitin-specific peptidase 48 to promote NHE3 degradation.
    Authors: Armando I, Villar V, Jones J, Lee H, Wang X, Asico L, Yu P, Yang J, Escano C, Pascua-Crusan A, Felder R, Jose P
    FASEB J, 2014;28(3):1422-34.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  34. A role for intersubunit interactions in maintaining SAGA deubiquitinating module structure and activity.
    Authors: Samara N, Ringel A, Wolberger C
    Structure, 2012;20(8):1414-24.
    Species: N/A
    Sample Types: Protein
    Applications: Bioassay
  35. Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15.
    Authors: Teyra J, Singer A, Schmitges F, Jaynes P, Kit Leng Lui S, Polyak M, Fodil N, Krieger J, Tong J, Schwerdtfeger C, Brasher B, Ceccarelli D, Moffat J, Sicheri F, Moran M, Gros P, Eichhorn P, Lenter M, Boehmelt G, Sidhu S
    Structure, 0;0(0):.
    Species: Human
    Sample Types: Transfected Whole Cells
    Applications: Bioassay

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Reviews for Recombinant Human Ubiquitin AMC Protein, CF

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Recombinant Human Ubiquitin AMC Protein, CF
By Anonymous on 06/01/2019
Application: Enzymatic activity in vitro
Reason for Rating: The substrates was able to be hydrolysed by USP7 and fluoresence was detected.