|Detection of Mouse CXCL9/MIG by Western Blot. Western blot shows lysates of mouse lung tissue. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Mouse CXCL9/MIG Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-492-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CXCL9/MIG at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Chemotaxis Induced by CXCL9/MIG and Neutralization by Mouse CXCL9/MIG Antibody. Recombinant Mouse CXCL9/MIG (Catalog #|
492-MM) 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 Mouse CXCL9/MIG (1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse CXCL9/MIG Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-492-NA). The ND50 is typically 6-20 µg/mL.
CXCL9, also known as MIG, is a member of the alpha subfamily of chemokines that lacks the ELR domain, and was initially identified as a lymphokine-activated gene in mouse macrophages. Human CXCL9 was subsequently cloned using mouse CXCL9 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 mouse CXCL9 cDNA encodes a 126 amino acid residue precursor protein with a 21 amino acid residue signal peptide that is cleaved to yield a 105 amino acid residue mature protein. CXCL9 has an extended carboxy-terminus containing greater than 50% basic amino acid 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.
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