Chemotaxis Induced by CX3CL1/Fractalkine and Neutralization by Rat CX3CL1/Fractalkine Antibody. |
Recombinant Rat CX3CL1/Fractalkine (Catalog # 536-FR) chemoattracts the BaF3 mouse pro‑B cell line transfected with human CX3CR1 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 Rat CX3CL1/Fractalkine (40 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat CX3CL1/Fractalkine Chemokine Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF537). The ND50 is typically 0.3-1.2 µg/mL.
CX3CL1/Fractalkine in Mouse Brain. |
CX3CL1/Fractalkine was detected in perfusion fixed frozen sections of mouse brain (cingulate cortex) using 1.7 µg/mL Goat Anti-Rat CX3CL1/Fractalkine Chemokine Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF537) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
CX3CL1, also named neurotactin, is a member of the delta chemokine subfamily that contains a novel C-X3-C motif. Unlike other known chemokines, CX3CL1 is a type 1 membrane protein containing a chemokine domain tethered on a long mucin-like stalk. Rat CX3CL1 cDNA encodes a 393 amino acid (aa) residue precursor protein with two alternative (21 aa or 24 aa residue) putative signal peptides, a 74 aa or 76 aa residue globular chemokine domain, a 238 aa residue stalk region rich in Gly, Pro, Ser and Thr and containing degenerate mucin-like repeats, a 19 aa residue transmembrane segment and a 36 aa residue cytoplasmic domain. The extracellular domain of CX3CL1 can potentially be released as a soluble protein by proteolysis at the conserved dibasic motif proximal to the transmembrane region. With the exception of the stalk region, rat CX3CL1 shares a high degree of amino acid sequence homology (83% sequence identity) with human and mouse CX3CL1. CX3CL1 is expressed in various tissues including heart, brain, lung, kidney, skeletal muscle, and testis. In rat brain, CX3CL1 expression was found to be localized principally to neurons. The expression of CX3CL1 was also reported to be up-regulated on activated endothelial cells. Membrane-bound CX3CL1 has been shown to promote adhesion of leukocytes. The soluble chemokine domain of human CX3CL1 was reported to be chemotactic for T cells and monocytes while the soluble chemokine domain of mouse CX3CL1 was reported to chemoattract neutrophils and T‑lymphocytes but not monocytes. CX3CR1, previously named V28 or chemokine beta receptor‑like 1, has been found to be a specific receptor for CX3CL1. In addition, US28, a 7TM receptor encoded by human cytomegalovirus that binds multiple CC chemokines, has also been shown to bind CX3CL1 with high-affinity.
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