Recombinant Human Ubiquitin AMC Protein, CF Summary
Product Specifications
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.
U-550
Formulation | Supplied as a solution in 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.
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Reconstitution Calculator
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.
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- Dang L.C., et al. (1998) Biochem. 37:1868-1879.
- Mason D.E., et al. (2004) Biochem 43:6535-6544.
- Stein R.L., et al. (1998) Biochem. 34:12616-12623.
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.
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Citations: Showing 1 - 10
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Targeting the Plasmodium falciparum UCHL3 ubiquitin hydrolase using chemically constrained peptides
Authors: King, HR;Bycroft, M;Nguyen, TB;Kelly, G;Vinogradov, AA;Rowling, PJE;Stott, K;Ascher, DB;Suga, H;Itzhaki, LS;Artavanis-Tsakonas, K;
Proceedings of the National Academy of Sciences of the United States of America
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay -
The BAP1 nuclear deubiquitinase is involved in the nonhomologous end-joining pathway of double-strand DNA repair through interaction with DNA-PK
Authors: Sato, H;Ito, T;Hayashi, T;Kitano, S;Erdjument-Bromage, H;Bott, MJ;Toyooka, S;Zauderer, M;Ladanyi, M;
Oncogene
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay -
Pharmaceutical targeting of OTUB2 sensitizes tumors to cytotoxic T cells via degradation of PD-L1
Authors: Ren, W;Xu, Z;Chang, Y;Ju, F;Wu, H;Liang, Z;Zhao, M;Wang, N;Lin, Y;Xu, C;Chen, S;Rao, Y;Lin, C;Yang, J;Liu, P;Zhang, J;Huang, C;Xia, N;
Nature communications
Species: Human
Sample Types: Phage Antibody Library
Applications: Bioassay -
Demonstrating the importance of porcine reproductive and respiratory syndrome virus papain-like protease 2 deubiquitinating activity in viral replication by structure-guided mutagenesis
Authors: Bailey-Elkin, BA;Knaap, RCM;De Silva, A;Boekhoud, IM;Mous, S;van Vught, N;Khajehpour, M;van den Born, E;Kikkert, M;Mark, BL;
PLoS pathogens
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay -
Altered ubiquitin signaling induces Alzheimer's disease-like hallmarks in a three-dimensional human neural cell culture model
Authors: Maniv, I;Sarji, M;Bdarneh, A;Feldman, A;Ankawa, R;Koren, E;Magid-Gold, I;Reis, N;Soteriou, D;Salomon-Zimri, S;Lavy, T;Kesselman, E;Koifman, N;Kurz, T;Kleifeld, O;Michaelson, D;van Leeuwen, FW;Verheijen, BM;Fuchs, Y;Glickman, MH;
Nature communications
Species: N/A
Sample Types: Recombinant Protein
Applications: Bioassay -
Accelerating inhibitor discovery for deubiquitinating enzymes
Authors: WC Chan, X Liu, RS Magin, NM Girardi, SB Ficarro, W Hu, MI Tarazona G, CA Starnbach, A Felix, G Adelmant, AC Varca, B Hu, AS Bratt, E DaSilva, NJ Schauer, I Jaen Maiso, EK Dolen, AX Ayala, JA Marto, SJ Buhrlage
Nature Communications, 2023-02-08;14(1):686.
Species: Human
Sample Types: Cell Lysates
Applications: Bioassay -
Targeting BAP1 with small compound inhibitor for colon cancer treatment
Authors: M Kang, SG Park, SA Lee, S Kim, D Lee, ME Shirbhate, SY Youn, KM Kim, SS Cha, J Kwon
Scientific Reports, 2023-02-08;13(1):2264.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Targeting USP10 induces degradation of oncogenic ANLN in esophageal squamous cell carcinoma
Authors: YF Cao, L Xie, BB Tong, MY Chu, WQ Shi, X Li, JZ He, SH Wang, ZY Wu, DX Deng, YQ Zheng, ZM Li, XE Xu, LD Liao, YW Cheng, LY Li, LY Xu, EM Li
Cell Death and Differentiation, 2022-12-16;0(0):.
Species: Human
Sample Types: Recombinant Proteins
Applications: Bioassay -
USP14-regulated allostery of the human proteasome by time-resolved cryo-EM
Authors: S Zhang, S Zou, D Yin, L Zhao, D Finley, Z Wu, Y Mao
Nature, 2022-04-27;605(7910):567-574.
Species: Human
Sample Types: Recombinant Proteins
Applications: Bioassay -
Repurposing the Pathogen Box compounds for identification of potent anti-malarials against blood stages of Plasmodium falciparum with PfUCHL3 inhibitory activity
Authors: H Bharti, A Singal, M Saini, PS Cheema, M Raza, S Kundu, A Nag
Scientific Reports, 2022-01-18;12(1):918.
Species: Human
Sample Types: Peptide
Applications: Bioassay -
A molecular sensor determines the ubiquitin substrate specificity of SARS-CoV-2 papain-like protease
Authors: S Patchett, Z Lv, W Rut, M Békés, M Drag, SK Olsen, TT Huang
Cell Reports, 2021-09-08;36(13):109754.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Small molecule inhibition of deubiquitinating enzyme JOSD1 as a novel targeted therapy for leukemias with mutant JAK2
Authors: J Yang, EL Weisberg, X Liu, RS Magin, WC Chan, B Hu, NJ Schauer, S Zhang, I Lamberto, L Doherty, C Meng, M Sattler, L Cabal-Hier, E Winer, R Stone, JA Marto, JD Griffin, SJ Buhrlage
Leukemia, 2021-07-29;0(0):.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Discovery of SARS-CoV-2 papain-like protease inhibitors through a combination of high-throughput screening and FlipGFP-based reporter assay
Authors: Z Xia, MD Sacco, C Ma, JA Townsend, N Kitamura, Y Hu, M Ba, T Szeto, X Zhang, X Meng, F Zhang, Y Xiang, MT Marty, Y Chen, J Wang
bioRxiv : the preprint server for biology, 2021-03-16;0(0):.
Species: Human
Sample Types: Protein
Applications: Bioassay -
The in-vitro effect of famotidine on sars-cov-2 proteases and virus replication
Authors: M Loffredo, H Lucero, DY Chen, A O'Connell, S Bergqvist, A Munawar, A Bandara, S De Graef, SD Weeks, F Douam, M Saeed, AH Munawar
Scientific Reports, 2021-03-08;11(1):5433.
Species: N/A
Sample Types: Protease
Applications: Bioassay -
Identification of ebselen and its analogues as potent covalent inhibitors of papain-like protease from SARS-CoV-2
Authors: E Weglarz-To, JM Tomczak, M Talma, M Burda-Grab, M Giurg, S Brul
Scientific Reports, 2021-02-11;11(1):3640.
Species: Human
Sample Types: Protein
Applications: Bioassay -
Typically inhibiting USP14 promotes autophagy in M1-like macrophages and alleviates CLP-induced sepsis
Authors: F Xu, Y Ma, W Huang, J Gao, M Guo, J Li, L Kong, G Liang, R Du, Q Xu, X Wu
Cell Death Dis, 2020-08-20;11(8):666.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
Turnip yellow mosaic virus protease binds ubiquitin suboptimally to fine-tune its deubiquitinase activity
Authors: S Fieulaine, MD Witte, CS Theile, M Ayach, HL Ploegh, I Jupin, S Bressanell
J. Biol. Chem., 2020-07-30;0(0):.
Species: Human, Virus
Sample Types: Recombinant Protein
Applications: Bioassay -
Selective USP7 inhibition elicits cancer cell killing through a p53-dependent mechanism
Authors: NJ Schauer, X Liu, RS Magin, LM Doherty, WC Chan, SB Ficarro, W Hu, RM Roberts, RE Iacob, B Stolte, AO Giacomelli, S Perera, K McKay, SA Boswell, EL Weisberg, A Ray, D Chauhan, S Dhe-Pagano, KC Anderson, JD Griffin, J Li, WC Hahn, PK Sorger, JR Engen, K Stegmaier, JA Marto, SJ Buhrlage
Sci Rep, 2020-03-24;10(1):5324.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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., 2019-10-28;15(10):e1008086.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
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, 2019-09-26;11(10):.
Species: Virus - PRRSV
Sample Types: Recombinant Protein
Applications: Bioassay -
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, 2019-07-23;10(4):.
Species: Virus
Sample Types: Protein
Applications: Bioassay -
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., 2019-03-26;0(0):.
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 -
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, 2019-01-31;0(0):.
Species: Human
Sample Types: Transfected Whole Cells
Applications: Bioassay -
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, 2019-01-27;0(0):1-20.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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, 2018-12-05;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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, 2018-05-18;9(38):24992-25007.
Applications: Bioassay -
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, 2018-03-28;9(1):1270.
Species: Human
Sample Types: Protein
Applications: Bioassay -
TRIM11 activates the proteasome and promotes overall protein degradation by regulating USP14
Authors: L Chen, G Zhu, EM Johns, X Yang
Nat Commun, 2018-03-26;9(1):1223.
Species: Human
Sample Types: Protein
Applications: Bioassay -
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, 2018-03-21;9(1):1162.
Species: Human
Sample Types: Protein
Applications: Bioassay -
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., 2018-02-17;0(0):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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., 2017-11-08;13(11):e1006714.
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 -
Ubiquitin Recognition of BAP1: Understanding its Enzymatic Function
Authors: P Hanpude, S Bhattachar, AK Singh, TK Maiti
Biosci. Rep., 2017-10-27;0(0):.
Applications: Enzyme Assay -
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., 2017-09-26;0(0):.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
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, 2017-06-15;8(6):e2877.
Applications: Enzyme Assay -
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., 2017-06-03;0(0):.
Applications: Bioassay -
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, 2017-03-28;6(0):.
Applications: Enzyme Assay -
A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibility
Authors: JF Beckmann, JA Ronau, M Hochstrass
Nat Microbiol, 2017-03-01;2(0):17007.
Applications: Bioassay -
Diverse mechanisms of metaeffector activity in an intracellular bacterial pathogen, Legionella pneumophila
Authors: Malene L Urbanus
Mol. Syst. Biol, 2016-12-16;12(12):893.
Applications: Bioassay -
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, 2016-08-12;113(35):E5192-201.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
Saturation scanning of ubiquitin variants reveals a common hot spot for binding to USP2 and USP21
Proc Natl Acad Sci USA, 2016-07-19;113(31):8705-10.
Species: Human
Sample Types: Recombinant Protein
Applications: Enzyme Assay -
BBX21, an Arabidopsis B-box protein, directly activates HY5 and is targeted by COP1 for 26S proteasome-mediated degradation
Proc Natl Acad Sci USA, 2016-06-20;113(27):7655-60.
Applications: Bioassay -
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, 2016-02-11;61(4):614-24.
Applications: Enzyme Assay -
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, 2015-08-19;290(42):25293-306.
Applications: Enzyme Assay -
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, 2015-01-15;10(1):e0115344.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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, 2014-04-11;20(12):3174-86.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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, 2014-03-15;454(0):247-53.
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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, 2013-12-05;28(3):1422-34.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
A role for intersubunit interactions in maintaining SAGA deubiquitinating module structure and activity.
Authors: Samara N, Ringel A, Wolberger C
Structure, 2012-07-05;20(8):1414-24.
Applications: Bioassay
FAQs
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Is Recombinant Human ISG15 Fluorescein Protein, CF (Catalog # UL-613) a suitable substrate for Recombinant His6-SARS Virus Papain-like Protease Protein, CF (Catalog # E-610)?
Recombinant His6-SARS Virus Papain-like Protease Protein, CF (Catalog # E-610) only cleaves at the C-terminus of ISG15 and/or Ubiquitin conjugates. Recombinant Human ISG15 Fluorescein Protein, CF (Catalog # UL-613) is not a suitable substrate because the fluorescein-labeled lysine residues are distributed throughout the protein, but not at the C-terminus. Suitable substrate candidates for catalog # E-610 include include Recombinant Human Ubiquitin AMC Protein, CF (Catalog #U-550), Recombinant Human Ubiquitin Rhodamine 110 Protein, CF (Catalog # U-555), and Recombinant Human ISG15 AMC Protein, CF (Catalog # UL-553).
Reviews for Recombinant Human Ubiquitin AMC Protein, CF
Average Rating: 4 (Based on 3 Reviews)
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Reason for Rating: A good pseudosubstrate for many DUBs
Reason for Rating: Works well but is expensive when working with DUBs with low activity in vitro.
Hydrolysis by USP8