R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.
ISG15 (22D2) Rabbit mAb #2758
Filter:
- WB
- IP
Western blot analysis of lysates from HeLa and A549 cells, untreated (-) or treated with IFN-α (1000 U/mL) (+) for 24 hours, using ISG15 (22D2) Rabbit mAb.
Supporting Data
| REACTIVITY | H |
| SENSITIVITY | Endogenous |
| MW (kDa) | 15 |
| Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
Species Cross-Reactivity Key:
- H-Human
- Related Products
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:25 |
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
Protocol
Specificity / Sensitivity
ISG15 (22D2) Rabbit mAb detects endogenous levels of the uncleaved precursor form of ISG15 protein. This antibody does not recognize the activated (cleaved) or conjugated forms of ISG15. The antibody does not cross-react with other ubiquitin family members, including ubiquitin, SUMO-1, SUMO-2, SUMO-3 and NEDD8.
Species Reactivity:
Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to amino acids at the carboxy terminus of human ISG15 protein.
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.
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