Technical datasheet
Description
The monoclonal antibody 6D4 recognizes human C-X-C motif chemokine 10 (IP-10), a protein of 98 amino acids. IP-10, also known as CXCL10, functions as ligand for the CXCR3 receptor. IP-10 belongs to the α-chemokine (C-X-C) family, which can be divided in two subfamilies: (1) potent chemoattractants for neutrophils, like IL-8 and (2) potent chemoattractants for lymphocytes, like the IFN inducible protein (IP)-10. IP-10 is produced by a wide variety of cell types ranging from neutrophils, dendritic cells and monocytes to hepatocytes, endothelial cells and keratinocytes. The cytokine is reported to be involved in a scala of inflammatory pathologies such as HIV, encephalitis, cutaneous T cell lymphoma, chronic hepatitis, psoriasis and acute anterior uveitis. Various observations strongly suggest a role for the C-X-C chemokines IL-8 and IP-10 in the regulation of angiogenic activity in cancer and in idiopathic pulmonary fibrosis. Furthermore IP-10 is associated with acute rejection processes estimated by the predictive properties of urinary IP-10 expression for the short- and long-term graft function after kidney transplantation.
Immunogen
Recombinant human IP-10
Formulation
0.5 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.
Species
Mouse IgG2a
Application
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F4
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FC3
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FS
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IA1,2
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IF4
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IP
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P
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W4
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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 notes
FC: Antibody 6D4 stains human CXCL10. For intracellular staining activated plasmacytoid dendritic cells were fixed with 2% paraformaldehyde and permeabilized with 0.5% saponin. All steps were performed in presence of 0.5 M EDTA to avoid cell aggregation (Ref.3)
W: Tissue was homogenized in loading sample buffer and subjected to SDS-PAGE. The band size is ~9 kDa (Ref.4).
IHC-F: Tissue sections were fixed 10 minutes in acetone and pretreated with avidin/biotin blocking kit. As positive control chronically inflamed liver was used and as negative control normal liver and isotype matched antibody (Ref.4).
Use
For immunohistochemistry, flow cytometry and Western blotting, 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 1:50.
Aliases
10 kDa interferon gamma-induced protein, CXCL10, Small-inducible cytokine B10
Positive control
Acitvated plasmacytoid dendritic cells
Negative control
Normal liver
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
References
1. Hamamdzic, D. et al; Reovirus triggers cell type-specific proinflammatory responses dependent on the autocrine action of IFNβ. Am J Physiol Lung Cell Mol Physiol 2001, 280: L18
2. Giustizieri, M et al; Nitric oxide donors suppress chemokine production by keratinocytes in vitro and in vivo. Am J Path 2002, 161: 1409
3. Bendriss-Vermare, N et al; Virus overrides the propensity of human CD40L-activated plasmacytoid dendritic cells to produce Th2 mediators through synergistic induction of IFN-gamma and Th1 chemokine production. J of Leukoc Biol. 2005, 78: 954
4. Curbishley, S et al; CXCR3 activation promotes lymphocyte transendothelial migration across human hepatic endothelium under fluid flow. Am J Pathol 2005, 167: 887
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 from the use or derivation of this product.
Also available
References
1. Hamamdzic, D. et al; Reovirus triggers cell type-specific proinflammatory responses dependent on the autocrine action of IFNβ. Am J Physiol Lung Cell Mol Physiol 2001, 280: L18
2. Giustizieri, M et al; Nitric oxide donors suppress chemokine production by keratinocytes in vitro and in vivo. Am J Path 2002, 161: 1409
3. Bendriss-Vermare, N et al; Virus overrides the propensity of human CD40L-activated plasmacytoid dendritic cells to produce Th2 mediators through synergistic induction of IFN-gamma and Th1 chemokine production. J of Leukoc Biol. 2005, 78: 954
4. Curbishley, S et al; CXCR3 activation promotes lymphocyte transendothelial migration across human hepatic endothelium under fluid flow. Am J Pathol 2005, 167: 887