Revision 1
#2743
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, FC-FP, E-P
Reactivity:
H M Mk
Sensitivity:
Endogenous
MW (kDa):
15
Source/Isotype:
Rabbit
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Flow Cytometry (Fixed/Permeabilized) | 1:200 |
Peptide ELISA (DELFIA) | 1:100 |
Storage
Specificity/Sensitivity
This antibody detects endogenous levels of both free and conjugated ISG15 protein. The antibody does not cross-react with other ubiquitin family members, including ubiquitin, SUMO1, SUMO2, SUMO3 and NEDD8.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids from human ISG15 protein. Antibodies are purified by protein A and peptide affinity chromatography.
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).
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting FC-FP: Flow Cytometry (Fixed/Permeabilized)
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 G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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Limited Uses
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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 1
#2743
ISG15 Antibody
Western blot analysis of lysates from HeLa, A549, RAW and COS cells, treated with or without IFN (1000 U/mL) for 24 hours, using ISG15 antibody.
Western blot analysis of NEDD8, Ubiquitin, ISG15 and SUMO-2/3 recombinant proteins (5 ng each), using NEDD8 (#2745) , Ubiquitin (#3936), ISG15 (#2743) and SUMO-2/3 (#4974) Antibodies.
Flow cytometric analysis of Hela cells, untreated (blue) or IFNa treated (green), using ISG15 antibody compared to a nonspecific negative control antibody (red).
Revision 1
#2743
ISG15 Antibody
The relationship between recombinant ISG15 protein concentration and assay optical density readings. Recombinant NEDD8 protein was used as a negative control.