Technical datasheet
Description
The monoclonal antibody F18 recognizes and neutralizes both natural and recombinant mouse alpha Interferon (IFN-α). IFN-α is a cytokine that belongs to the type I interferons (IFN-I). IFN-α is secreted by many cell types including lymphocytes (NK cells, B-cells and T-cells), macrophages, fibroblasts, endothelial cells, osteoblasts, microglia and others. Interferons stimulate both macrophages and NK cells to elicit an anti-viral response, and are also active against tumors. Although all cells can produce IFN-I, plasmacytoid dendritic cells (pDCs) produce 1,000-fold higher levels than other cell types, and are responsible for systemic IFN-I responses to many viruses. They are coined as the natural IFN-producing cells. However, under deprived pDC condition, other dendritic cells are capable of producing high levels of IFN-I.
Interferons were initially characterized for their ability to ‘interfere' with viral replication, slow cell proliferation, and profoundly alter immunity. IFN-α has several regulatory roles and diverse biological activities, including control of cellular and humoral immune responses, inflammation, and tumor regression. In addition, IFN-α participates in the regulation of various cellular and humoral processes such as the endocrine system modulates behavior, brain activity, temperature, glucose sensitive neurons, feeding pattern and opiate activity.
With the availability of monoclonal antibodies directed against IFN-α, it is possible to interpret results obtained from crude materials containing both IFN-α and IFN-β. The difficulties in studying in vitro and in vivo effects of ‘type 1’ Interferons arise from the fact that both alpha and beta Interferons are produced in response to the same stimuli and also seem to act via the same receptor. These Interferon activities can only be distinguished from one another by use of specific neutralizing antibodies.
Cross Reactivity
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Cross reactant
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Reactivity
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Human IFN
Mouse IFN-β
Mouse IFN-g
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No
No
No
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Formulation
0.5 mg (100 µg/ml) 0.2 µm filtered antibody solution in PBS (exact concentration is indicated on the label)
Species
Rat IgG1
Application
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F
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FC2
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FS3
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IA1
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IF
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IP
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P
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W
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Yes
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No
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N.D.
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application IA has been tested by Hycult Biotech. Application IP is based on personal communication.
Application notes
FC: For intracellular staining of IFN-α, cells can be fixed in 1 % formaldehyde; blocked and permeabilized in 0.2 % saponin, 5 % normal rabbit serum for 30 minutes on ice. (Ref.2)
FS: Neutralization of IFN-α by adding 1 µg antibody F18 per mouse i.v. before treatment with 35 µg LPS i.p., decreased the LPS-induced IL-1β serum response. (Ref.3).
Use
For flow cytometry dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:10. Before use in biological assays, the product must be filter sterilized and depending on the concentration to be used dialyzed against culture medium to remove the sodium azide added. For neutralisation of biological activity dilutions have to be made according to the amount of Interferon to be inactivated.
Aliases
IFN-α
Positive control
Plasmacytoid dendritic cells
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.
References
1. Dalod, M et al; Interferon α/β and interleukin 12 responses to viral infections: pathways regulating dendritic cell cytokine expression in vivo. J Exp Med 2002, 195: 517
2. Diebold, S et al; Viral infection switches nonplasmacytoid dendritic cells into high interferon producers. Nature 2003, 424: 324
3. Joshi, V et al; A role for Stat1 in the regulation of lipopolysaccharide-induced Interleukin-1β expression. J Interferon Cytokine Res 2006, 26: 739.
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Also available
References
1. Dalod, M et al; Interferon α/β and interleukin 12 responses to viral infections: pathways regulating dendritic cell cytokine expression in vivo. J Exp Med 2002, 195: 517
2. Diebold, S et al; Viral infection switches nonplasmacytoid dendritic cells into high interferon producers. Nature 2003, 424: 324
3. Joshi, V et al; A role for Stat1 in the regulation of lipopolysaccharide-induced Interleukin-1β expression. J Interferon Cytokine Res 2006, 26: 739.