Recombinant Human ISG15 AMC Protein, CF

Catalog # Availability Size / Price Qty
UL-553-050
Product Details
Citations (9)
FAQs
Reviews

Recombinant Human ISG15 AMC Protein, CF Summary

Activity
Recombinant Human ISG15 AMC is a fluorogenic substrate for some ISG15-specific isopeptidases. Release of AMC fluorescence can be monitored with an excitation wavelength of 380 nm and an emission wavelength of 460 nm. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human ISG15 AMC concentration of 0.1-1 μM.
Source
E. coli-derived human ISG15/UCRP protein
Contains underivatized and C-terminal AMC derivatized protein, quantity is by derivatized content
Accession #
Predicted Molecular Mass
19 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.

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.
  • 12 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
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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.

References
  1. Yuan, W. & R.M. Krug (2001) EMBO J. 20:362.
  2. Zhao, C. et al. (2004) Proc. Natl. Acad. Sci. USA 101:7578.
  3. Zou, W. & D.E. Zhang (2006) J. Biol. Chem. 281:3989.
  4. Wong, J.J. et al. (2006) Proc. Natl. Acad. Sci. USA 103:10735.
  5. Malakhov, M.P. et al. (2002) J. Biol. Chem. 277:9976.
  6. Zhang, D. & D.-E. Zhang (2011) J. Interferon Cytokine Res. 31:119.
  7. Jeon, Y.J. et al. (2012) J. Clin. Invest. 122:2622.
  8. Harty, R.N. et al. (2009) J. Innate. Immun. 1:397.
  9. Owashi, M. et al. (2003) Biochem. Biophys. Res. Commun. 309:533.
  10. Bogunovic, D. et al. (2012) Science 337:1684.
Long Name
ISG15 Ubiquitin-like Modifier
Entrez Gene IDs
9636 (Human); 100038882 (Mouse); 298693 (Rat)
Alternate Names
G1P2; G1P2interferon-induced 17-kDa/15-kDa protein; IFI15; IFI15hUCRP; interferon, alpha-inducible protein (clone IFI-15K); Interferon-induced 15 kDa protein; Interferon-induced 17 kDa protein; interferon-stimulated protein, 15 kDa; IP17; ISG15 ubiquitin-like modifier; ISG15; Ubiquitin cross-reactive protein; ubiquitin-like protein ISG15; UCRP; UCRPIP17

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.

9 Citations: Showing 1 - 9
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  1. 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
  2. A family of unconventional deubiquitinases with modular chain specificity determinants
    Authors: T Hermanns, C Pichlo, I Woiwode, K Klopffleis, KF Witting, H Ovaa, U Baumann, K Hofmann
    Nat Commun, 2018;9(1):799.
    Applications: Bioassay
  3. Structural insights into the interaction of coronavirus papain-like proteases and interferon-stimulated gene product 15 from different species
    Authors: CM Daczkowski, JV Dzimianski, JR Clasman, O Goodwin, AD Mesecar, SD Pegan
    J. Mol. Biol., 2017;0(0):.
    Applications: Bioassay
  4. X-ray Structure and Enzymatic Activity Profile of a Core Papain-like Protease of MERS Coronavirus with utility for structure-based drug design
    Authors: JR Clasman, YM Báez-Santo, RC Mettelman, A O'Brien, SC Baker, AD Mesecar
    Sci Rep, 2017;7(0):40292.
    Species: N/A
    Sample Types: Protein
    Applications: Enzyme Assay
  5. Structural insights into the interaction of coronavirus papain-like proteases and interferon-stimulated gene product 15 from different species
    Authors: CM Daczkowski, JV Dzimianski, JR Clasman, O Goodwin, AD Mesecar, SD Pegan
    J. Mol. Biol., 2017;0(0):.
    Applications: Bioassay
  6. 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
  7. 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
  8. 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.
  9. High yield expression of catalytically active USP18 (UBP43) using a Trigger Factor fusion system.
    Authors: Basters A, Ketscher L, Deuerling E, Arkona C, Rademann J, Knobeloch K, Fritz G
    BMC Biotechnol, 2012;12(0):56.
    Species: N/A
    Sample Types: Recombinant Protein
    Applications: Bioassay

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