Intracellular Staining by Flow Cytometry
|Detection of CXCL9/MIG in THP‑1 Human Cell Line by Flow Cytometry. THP‑1 human acute monocytic leukemia cell line treated with Recombinant Human IFN‑ gamma (Catalog # 285-IF) in the presence of Monensin for 24 hours was stained with Mouse Anti-Human CXCL9/MIG Fluorescein‑conjugated Monoclonal Antibody (Catalog # IC392F, filled histogram) or isotype control antibody (Catalog # IC002F, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.|
CXCL9, a member of the alpha subfamily of chemokines that lack the ELR domain, was initially identified as a lymphokine-activated gene in mouse macrophages. Human CXCL9 was subsequently cloned using mouse MIG cDNA as a probe. The CXCL9 gene is induced in macrophages and in primary glial cells of the central nervous system specifically in response to IFN-gamma. CXCL9 has been shown to be a chemoattractant for activated T-lymphocytes and TIL but not for neutrophils or monocytes. The human CXCL9 cDNA encodes a 125 amino acid (aa) residue precursor protein with a 22 aa residue signal peptide that is cleaved to yield a 103 aa residue mature protein. CXCL9 has an extended carboxy-terminus containing greater than 50% basic aa residues and is larger than most other chemokines. The carboxy-terminal residues of CXCL9 are prone to proteolytic cleavage resulting in size heterogeneity of natural and recombinant CXCL9. CXCL9 with large carboxy-terminal deletions have been shown to have diminished activity in the calcium flux assay. A chemokine receptor (CXCR3) specific for CXCL9 and IP-10 has been cloned and shown to be highly expressed in IL-2-activated T-lymphocytes. The E. coli-expressed CXCL9 preparations produced at R&D Systems have been shown to contain greater than 80% full length CXCL9.
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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