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Recombinant Human ISG15 Fluorescein Protein, CF

Catalog #: UL-613 Datasheet / COA / SDS

Discontinued Product

UL-613 has been discontinued.
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Recombinant Human ISG15 Fluorescein Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain
Activity
Recombinant Human ISG15 Fluorescein is ideal for use in assays requiring fluorescent detection. Optimal fluorescence at pH 8.0 is monitored with an excitation wavelength of 490 nm and an emission wavelength of 515 nm. Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human ISG15 Fluorescein concentration of 0.1-1 μM.
Source
E. coli-derived human ISG15/UCRP protein
Accession #
P05161
Predicted Molecular Mass
18 kDa (unlabeled), extent of labeling varies by lot

Product Datasheets

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UL-613

Product Datasheet
COA
SDS

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-613

Formulation

X mg/ml (XμM) in 50 mM HEPES, 100 mM NaCl, 1 mM DTT and 10% (v/v) glycerol, pH 7.5
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.
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

 

ISG15 modified with fluorescein via primary amine coupling, resulting in the modification of lysine residues as well as the N-terminus. This labeled ISG15 allows for direct detection spectophotometrically with higher efficiency and sensitivity than with antibodies.

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.
  11. Dao C.T. and Zhang D.E. (2005) Front. Biosci. 10:2346-2365.
  12. Dao C.T. and Zhang D.E. (2005) Front. Biosci. 10:2701-2722.
  13. D'Cunha J., et al. (1996) Proc. Natl. Acad. Sci. 93:211-215.
  14. Kim K.I. and Zhang D. (2003) Biochem. Biophys. Res. Comm. 307:431-434.
  15. Owhashi M., et al. (2003) Biochem. Biophys. Res. Comm. 309:533-539.
  16. Narasimhan J., et al. (2005) J. Biol Chem. 280:27356-27365.
  17. Potter J.L., et al. (1999) J. Biol Chem. 267:25061068.
  18. Ritchie K.J. and Zhang D.E. (2004) Sem. Cell. Dev. Biol. 2: 237-246.        
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

FAQs

  1. 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).

  2. Is Recombinant Human ISG15 Fluorescein Protein, CF (Catalog # UL-613) a suitable substrate for Recombinant SARS-CoV-2 GST-Papain-like Protease, CF (Catalog # E-611)?

    • Recombinant SARS-CoV-2 GST-Papain-like Protease, CF (Catalog # E-611) 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-611 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).

  3. Is Recombinant Human ISG15 Fluorescein Protein, CF (Catalog # UL-613) a suitable substrate for Recombinant MERS-CoV His6 Papain-like Protease Protein, CF (Catalog # E-609)?

    • Recombinant MERS-CoV His6 Papain-like Protease Protein, CF (Catalog # E-609) 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 Catago #E-609 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).

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