Recombinant Human ISG15 AMC Protein, CF Summary
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.
UL-553
| Formulation | X mg/ml (X µM) in 50 mM HEPES pH 7.5, 100 mM NaCl, 20% Glycerol (v/v), 2mM DTT |
| 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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Background: ISG15/UCRP
Interferon-stimulated Gene 15 (ISG15), also known as Ubiquitin Cross-reacting Protein (UCRP), is a Ubiquitin-like protein that is covalently coupled to target proteins in a process termed ISGylation. It is a 165 amino acid (aa) polypeptide with a predicted molecular weight of 18 kDa. ISG15/UCRP exhibits 66% aa sequence identity with its mouse ortholog. Structurally, ISG15/UCRP consists of two tandem Ubiquitin-like domains that share a similar 3-dimensional structure with Ubiquitin and other Ubiquitin-like modifiers including NEDD8 and SUMO1. Modification of targets by ISG15/UCRP occurs in a stepwise enzymatic process similar to that of Ubiquitin. Enzymes regulating ISGylation include the activating (E1) enzyme UBE1L, the conjugating (E2) enzyme UbcH8, and ligases (E3) such as EFP/TRIM25 and HERC5 (1-4). Removal of ISG15/UCRP is catalyzed by the deconjugating enzyme UBP43/USP18 (5). Functionally, ISG15/UCRP has putative roles in the immune response and tumorigenesis. This is reflected by intracellular ISG15/UCRP targets that include Cyclin D1, tumor suppressor p63, IRF3, and a range of viral proteins (6-8). It is induced by type 1 interferons and microbial infection, and knockout mice exhibit an increased sensitivity to infection by some viruses (6). ISG15/UCRP can also be secreted by cells of the immune system and may act in a cytokine-like manner (9). For instance, it is produced by human granulocytes in response to mycobacterium exposure, and natural killer cells and T cells respond to extracellular ISG15/UCRP with IFN-gamma production (10). Further supporting a role in immune function, ISG15/UCRP mutations are associated with MSMD, an inherited disorder characterized by increased susceptibility to mycobacterial infection (10).
This fluorogenic substrate for ISG15 hydrolases is based on the carboxy-terminus derivatization of ISG15 with 7-amido-4-methylcoumarin (AMC). ISG15 AMC is useful for studying enzymes (such as UBP43 and Papain-Like Protease from SARS coronavirus) when detection sensitivity or continuous monitoring of activity is essential.
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- Zhao, C. et al. (2004) Proc. Natl. Acad. Sci. USA 101:7578.
- Zou, W. & D.E. Zhang (2006) J. Biol. Chem. 281:3989.
- Wong, J.J. et al. (2006) Proc. Natl. Acad. Sci. USA 103:10735.
- Malakhov, M.P. et al. (2002) J. Biol. Chem. 277:9976.
- Zhang, D. & D.-E. Zhang (2011) J. Interferon Cytokine Res. 31:119.
- Jeon, Y.J. et al. (2012) J. Clin. Invest. 122:2622.
- Harty, R.N. et al. (2009) J. Innate. Immun. 1:397.
- Owashi, M. et al. (2003) Biochem. Biophys. Res. Commun. 309:533.
- Bogunovic, D. et al. (2012) Science 337:1684.
Citations for Recombinant Human ISG15 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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- Crystal structure and activity-based labeling reveal the mechanisms for linkage-specific substrate recognition by deubiquitinase USP9XAuthors: P Paudel, Q Zhang, C Leung, HC Greenberg, Y Guo, YH Chern, A Dong, Y Li, M Vedadi, Z Zhuang, Y TongProc. Natl. Acad. Sci. U.S.A., 2019;0(0):.Applications: Bioassay
- A family of unconventional deubiquitinases with modular chain specificity determinantsAuthors: T Hermanns, C Pichlo, I Woiwode, K Klopffleis, KF Witting, H Ovaa, U Baumann, K HofmannNat Commun, 2018;9(1):799.Applications: Bioassay
- Structural insights into the interaction of coronavirus papain-like proteases and interferon-stimulated gene product 15 from different speciesAuthors: CM Daczkowski, JV Dzimianski, JR Clasman, O Goodwin, AD Mesecar, SD PeganJ. Mol. Biol., 2017;0(0):.Applications: Bioassay
- X-ray Structure and Enzymatic Activity Profile of a Core Papain-like Protease of MERS Coronavirus with utility for structure-based drug designAuthors: JR Clasman, YM Báez-Santo, RC Mettelman, A O"Brien, SC Baker, AD MesecarSci Rep, 2017;7(0):40292.Species: N/ASample Types: ProteinApplications: Enzyme Assay
- Structural insights into the interaction of coronavirus papain-like proteases and interferon-stimulated gene product 15 from different speciesAuthors: CM Daczkowski, JV Dzimianski, JR Clasman, O Goodwin, AD Mesecar, SD PeganJ. Mol. Biol., 2017;0(0):.Applications: Bioassay
- A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibilityAuthors: JF Beckmann, JA Ronau, M HochstrassNat Microbiol, 2017;2(0):17007.Species: N/ASample Types: ProteinApplications: Bioassay
- X-ray Structural and Functional Studies of the Three Tandemly Linked Domains ofNon-structural Protein 3 (nsp3) from Murine Hepatitis Virus Reveal ConservedFunctions.Authors: Chen Y, Savinov S, Mielech A, Cao T, Baker S, Mesecar AJ Biol Chem, 2015;290(42):25293-306.Species: N/ASample Types: ProteinApplications: Enzyme Assay
- The vOTU domain of highly-pathogenic porcine reproductive and respiratorysyndrome virus displays a differential substrate preference.Authors: Deaton M, Spear A, Faaberg K, Pegan SVirology, 2014;454(0):247-53.
- High yield expression of catalytically active USP18 (UBP43) using a TriggerFactor fusion system.Authors: Basters A, Ketscher L, Deuerling E, Arkona C, Rademann J, Knobeloch K, Fritz GBMC Biotechnol, 2012;12(0):56.Species: N/ASample Types: Recombinant ProteinApplications: Bioassay
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