#P05161
9636
Product Information
Product Usage Information
CST recommends transfection with 100 nM SignalSilence® ISG15 siRNA II 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.
Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.
Storage
Specificity / Sensitivity
Species Reactivity:
Human
Product Description
Background
Interferon-stimulated 15 kDa protein (ISG15), also known as ubiquitin cross-reactive protein (UCRP), is a member of the ubiquitin-like protein family and functions in various biological pathways from pregnancy to innate immune responses (1). Expression of ISG15 is stimulated by cellular exposure to type 1 interferons α and β, in addition to infection with viruses such as influenza B (2,3). After exposure to type I interferons, both lymphocytes and monocytes, in addition to some fibroblasts and epithelial cells, release ISG15 into culture medium (1,4). ISG15 has been shown to function as a cytokine, stimulating interferon γ secretion by monocytes and macrophages, proliferation of natural killer cells, and chemotactic responses in neutrophils (4,5). ISG15 has also been shown to function intracellularly, being covalently conjugated to other proteins by E1 (Ube1L), E2 (UbcH8) and E3 ligases via a multi-step process analogous to ubiquitination (6,7). ISG15 is removed from proteins by the ubiquitin processing protease Ubp43 (8). ISG15-protein conjugation (ISGylation) is induced by type 1 interferons, and target proteins include the serine protease inhibitor Serpin 2A, PLCγ1, ERK1/2, Jak1 and Stat1 (9,10). Unlike ubiquitination, ISGylation does not target proteins for degradation, rather ISGylation increases Jak1 and Stat1 activity, enhancing the cellular response to interferons (11).
- Ritchie, K.J. and Zhang, D.E. (2004) Semin. Cell Dev. Biol. 15, 237-246.
- Korant, B.D. et al. (1984) J. Biol. Chem. 259, 14835-14839.
- Haas, A.L. et al. (1987) J. Biol. Chem. 262, 11315-11323.
- Knight, E. and Cordova, B. (1991) J. Immunol. 146, 2280-2284.
- D'Cunha, J. et al. (1996) Proc. Natl. Acad. Sci. USA 93, 211-215.
- Loeb, K.R. and Haas, A.L. (1992) J. Biol. Chem. 267, 7806-7813.
- Zhao, C. et al. (2005) Proc. Natl. Acad. Sci. USA 102, 10200-10205.
- Malakhov, M.P. et al. (2002) J. Biol. Chem. 277, 9976-9981.
- Malakhov, M.P. et al. (2003) J. Biol. Chem. 278, 16608-16613.
- Hamerman, J.A. et al. (2002) J. Immunol. 168, 2415-2423.
- Malakhova, O.A. et al. (2003) Genes Dev. 17, 455-460.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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