Recombinant Human ISG15 Protein, Low Endotoxin, CF
Recombinant Human ISG15 Protein, Low Endotoxin, CF Summary
Product Specifications
Recombinant Human ISG15 can be conjugated to substrate proteins via the subsequent actions of an ISG15-activating (E1) enzyme, an ISG15-conjugating (E2) enzyme, and an ISG15 ligase (E3). Reaction conditions will need to be optimized for each specific application. We recommend an initial Recombinant Human ISG15 concentration of 1-10 μM for conjugation reactions. For cell-based applications, optimal concentrations will need to be established by the investigator.
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-605
| Formulation | X mg/ml (X μM) in 50 mM HEPES pH 7.5, 100 mM NaCl, 5% (v/v) Glycerol, 2mM TCEP |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
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. The mature form of ISG15 is a 156 amino acid (aa) polypeptide with a predicted molecular weight of 17 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).
- Yuan, W. & R.M. Krug (2001) EMBO J. 20:362.
- 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.
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