CX3CL1/Fractalkine Antibody [Unconjugated]

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.

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.

• Reactivity: Rt
• Applications: WB, IHC, B/N, ELISA(Cap)
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

CX3CL1/Fractalkine Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant rat CX3CL1/Fractalkine
  Gln25-Gly100
  Accession # O55145
• Specificity: Detects rat CX3CL1/Fractalkine in ELISAs and Western blots. Neutralizes the biological activity of recombinant human CX3CL1 and recombinant mouse CX3CL1 with similar effectiveness.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CX3CL1
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • ELISA Capture (Matched Antibody Pair) 0.2-0.8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • Neutralization 0.3-1.2 ug/mL
  • ELISA Standard (Matched Pair)
• Application Notes: ELISA Capture: Rat CX3CL1/Fractalkine Chemokine Domain Antibody (Catalog # AF537)
  ELISA Detection:Rat CX3CL1/Fractalkine Chemokine Domain Biotinylated Antibody (Catalog # BAF537)
  Standard: Recombinant Rat CX3CL1/Fractalkine (Catalog # 536-FR)

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CX3CL1/Fractalkine Antibody [Unconjugated]

• ABCD-3
• C3Xkine
• chemokine (C-X3-C motif) ligand 1
• CX3C membrane-anchored chemokine
• CX3CL1
• CXC3
• CXC3C
• FKN
• Fractalkine
• Neurotactin
• neurotactin)
• NTNsmall inducible cytokine subfamily D (Cys-X3-Cys), member 1 (fractalkine
• NTTSmall-inducible cytokine D1
• SCYD1C-X3-C motif chemokine 1
• small inducible cytokine subfamily D (Cys-X3-Cys), member-1

Background

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.

1. Kledal, T.N. et al. (1998) FEBS Lett. 441:209.
2. Combadiere, C. et al. (1998) J. Biol. Chem. 273:23799.
3. Harrison, J.L. et al. (1998) Proc. Natl. Acad. Sci. USA 95:10896.
4. Rossi, D.L. et al. (1998) Genomics 47:163.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CX3CL1/Fractalkine Antibody (AF537)(6)

Publications using AF537

• S Castro-Sán, ÁJ García-Yag, T López-Royo, M Casarejos, JL Lanciego, I Lastres-Be Cx3cr1-deficiency exacerbates alpha-synuclein-A53T induced neuroinflammation and neurodegeneration in a mouse model of Parkinson&#039;s disease Glia, 2018;0(0):. 2018 [PMID: 29624735] (IHC, Mouse)
• José L M Madrigal Noradrenaline induces CX3CL1 production and release by neurons Neuropharmacology, 2017;114(0):146-155. 2017 [PMID: 27923568] (Western Blot, Rat)
• Yeh P, Huang H, Yang C, Yang W, Yang C Astaxanthin Inhibits Expression of Retinal Oxidative Stress and Inflammatory Mediators in Streptozotocin-Induced Diabetic Rats. PLoS ONE, 2016;11(1):e0146438. 2016 [PMID: 26765843] (Western Blot, Rat)
• Clark A, Gruber-Schoffnegger D, Drdla-Schutting R, Gerhold K, Malcangio M, Sandkuhler J Selective activation of microglia facilitates synaptic strength. J Neurosci, 2015;35(11):4552-70. 2015 [PMID: 25788673] (Neutralization, Rat)
• Cook A, Hippensteel R, Shimizu S, Nicolai J, Fatatis A, Meucci O Interactions between chemokines: regulation of fractalkine/CX3CL1 homeostasis by SDF/CXCL12 in cortical neurons. J. Biol. Chem., 2010;285(14):10563-71. 2010 [PMID: 20124406] (Western Blot, Rat)
• Clark AK, Yip PK, Grist J, Gentry C, Staniland AA, Marchand F, Dehvari M, Wotherspoon G, Winter J, Ullah J, Bevan S, Malcangio M Inhibition of spinal microglial cathepsin S for the reversal of neuropathic pain. Proc. Natl. Acad. Sci. U.S.A., 2007;104(25):10655-60. 2007 [PMID: 17551020] (Neutralization, Rat)

Bioinformatics

• Gene Symbol: CX3CL1
• Uniprot:
  • O55145()