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.
Flow cytometry, Frozen sections, Immuno assays, Immuno fluorescence, Western blot
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).
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.
Acitvated plasmacytoid dendritic cells