Rat CX3CL1/Fractalkine Chemokine Domain Antibody

(4 citations)
(2 Reviews)
  
  • Species Reactivity
    Rat
  • Specificity
    Detects rat CX3CL1/Fractalkine in ELISAs and Western blots. In Western blots, less than 5% cross-reactivity with recombinant human (rh) Eotaxin is observed. Neutralizes the biological activity of rhCX3CL1 and rmCX3CL1 with similar effectiveness.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant rat CX3CL1/Fractalkine
    Gln25-Gly100
    Accession # O55145
  • Formulation
    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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Rat CX3CL1/Fractalkine (Catalog # 536-FR)
  • Immunohistochemistry
    5-15 µg/mL
    See below
    • Rat CX3CL1/Fractalkine Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Rat CX3CL1/Fractalkine Chemokine Domain Antibody (Catalog # AF537)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Rat CX3CL1/Fractalkine Chemokine Domain Biotinylated Antibody (Catalog # BAF537)
  • ELISA Standard
     
    Recombinant Rat CX3CL1/Fractalkine Protein (Catalog # 536-FR)
  • Neutralization
    Measured by its ability to neutralize CX3CL1/Fractalkine-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with human CX3CR1. The Neutralization Dose (ND50) is typically 0.3-1.2 µg/mL in the presence of 40 ng/mL Recombinant Rat CX3CL1/Fractalkine.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.

Immunohistochemistry
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.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: CX3CL1/Fractalkine

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.

  • References:
    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.
  • Entrez Gene IDs:
    6376 (Human); 20312 (Mouse); 89808 (Rat)
  • Alternate Names:
    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
Related Research Areas
Citations:

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.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Astaxanthin Inhibits Expression of Retinal Oxidative Stress and Inflammatory Mediators in Streptozotocin-Induced Diabetic Rats.
    Authors: Yeh P, Huang H, Yang C, Yang W, Yang C
    PLoS ONE, 2016;11(1):e0146438.
    Species: Rat
    Sample Type: Cell Lysates
    Application: WB
  2. Selective activation of microglia facilitates synaptic strength.
    Authors: Clark A, Gruber-Schoffnegger D, Drdla-Schutting R, Gerhold K, Malcangio M, Sandkuhler J
    J Neurosci, 2015;35(11):4552-70.
    Species: Rat
    Sample Type: Whole Cells
    Application: Neut
  3. Interactions between chemokines: regulation of fractalkine/CX3CL1 homeostasis by SDF/CXCL12 in cortical neurons.
    Authors: Cook A, Hippensteel R, Shimizu S, Nicolai J, Fatatis A, Meucci O
    J. Biol. Chem., 2010;285(14):10563-71.
    Species: Rat
    Sample Type: Cell Lysates
    Application: WB
  4. Inhibition of spinal microglial cathepsin S for the reversal of neuropathic pain.
    Authors: Clark AK, Yip PK, Grist J, Gentry C, Staniland AA, Marchand F, Dehvari M, Wotherspoon G, Winter J, Ullah J, Bevan S, Malcangio M
    Proc. Natl. Acad. Sci. U.S.A., 2007;104(25):10655-60.
    Species: Rat
    Sample Type: In Vivo
    Application: In vivo
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Normal Goat IgG Control

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Average Rating: 4 (Based on 2 reviews)

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We have 2 reviews tested in 2 applications: Immunocytochemistry/Immunofluorescence, Immunohistochemistry.
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Images Ratings Applications Species Reviewed By Date Details
Immunocytochemistry/Immunofluorescence Rat CX3CL1/Fractalkine Chemokine Domain Antibody AF537
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 ICC/IF Anonymous 12/19/2016
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Immunocytochemistry/Immunofluorescence Rat CX3CL1/Fractalkine Chemokine Domain Antibody AF537
Immunocytochemistry/Immunofluorescence: Rat CX3CL1/Fractalkine Chemokine Domain Antibody [AF537]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedCortical neurons

Other Experimental Details

Other Experimental DetailsResults published in:Article title: Noradrenaline induces CX3CL1 production and release by neuronsArticle reference: NP6524Journal title: NeuropharmacologyFinal version published online: 08-Dec-2016DOI information: 10.1016/j.neuropharm.2016.12.001
Immunohistochemistry Rat CX3CL1/Fractalkine Chemokine Domain Antibody AF537
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 IHC Anonymous 12/01/2016
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Immunohistochemistry Rat CX3CL1/Fractalkine Chemokine Domain Antibody AF537
Immunohistochemistry: Rat CX3CL1/Fractalkine Chemokine Domain Antibody [AF537]

Summary

ApplicationImmunohistochemistry
Sample TestedBrain (cortex) tissue

Other Experimental Details

Other Experimental DetailsTraumatic brain injury site in rat brainFractalkine antibody (red) and DAPI (blue)

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