|CXCL9/MIG in THP-1 Human Cell Line. CXCL9/MIG was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line stimulated with IFN-gamma for 24 hours using Mouse Anti-Human CXCL9/MIG Monoclonal Antibody (Catalog # MAB392) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
|Chemotaxis Induced by CXCL9/MIG and Neutralization by Human CXCL9/MIG Antibody. Recombinant Human CXCL9/MIG (Catalog # 392-MG) chemoattracts the BaF3 mouse pro‑B cell line transfected with mouse CXCR3 in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Human CXCL9/MIG (0.25 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human CXCL9/MIG Monoclonal Antibody (Catalog # MAB392). The ND50 is typically 4-20 µg/mL.|
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.
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ELISA: Human CXCL9/MIG Antibody [MAB392]
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