Rat CX3CL1/Fractalkine Chemokine Domain Antibody

R&D Systems | Catalog # AF537

R&D Systems

Key Product Details

Species Reactivity

Validated:

Rat

Cited:

Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Immunocytochemistry, In vivo assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CX3CL1/Fractalkine Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Rat CX3CL1/Fractalkine Chemokine Domain Antibody

Chemotaxis Induced by CX3CL1/Fractalkine and Neutralization by Rat CX3CL1/Fractalkine Antibody.

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 antibody in Mouse Brain by Immunohistochemistry (IHC-Fr).

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.
Detection of Rat CX3CL1/Fractalkine by Immunohistochemistry

Detection of Rat CX3CL1/Fractalkine by Immunohistochemistry

Upregulation of the expression of CX3CL1 and CX3CR1 in the hippocampus of 2VO and miR-195 loss-of-function rats. a, b Expression of CX3CL1 (red) and CX3CR1 (green) in the hippocampus of sham and 2VO rats was shown by immunofluorescence staining. The scale bar is 500 um. c Expression of CX3CL1 and CX3CR1 in the hippocampus of sham and 2VO rats detected by western blot technique. Bars represent the mean ± SD, n = 6, *P < 0.05 vs sham group, Student’s t test. d Lenti-pre-AMO-miR-195 upregulated the expression of CX3CL1 in rat hippocampus. Mean ± SD, n = 6, *P < 0.05 vs sham; #P < 0.05 vs lenti-pre-AMO-miR-195. Data were analyzed using one-way ANOVA followed by Tukey test. e Lenti-pre-AMO-miR-195 upregulated the expression of CX3CR1 in the rat hippocampus. Mean ± SD, n = 6, *P < 0.05 vs sham; #P < 0.05 vs lenti-pre-AMO-miR-195. Data were analyzed using one-way ANOVA followed by Tukey test Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/32819407), licensed under a CC-BY license. Not internally tested by R&D Systems.
Rat CX3CL1 / Fractalkine ELISA Standard Curve

Rat CX3CL1 / Fractalkine ELISA Standard Curve

Recombinant Rat CX3CL1/Fractalkine (Catalog # 536-FR) was serially diluted and captured by Goat Anti-Rat CX3CL1/Fractalkine Chemokine Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF537) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Rat CX3CL1/Fractalkine Chemokine Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF537) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Rat CX3CL1 / Fractalkine ELISA Standard Curve

Rat CX3CL1 / Fractalkine ELISA Standard Curve

Recombinant Rat CX3CL1/Fractalkine (Catalog # 536-FR) was serially diluted and captured by Goat Anti-Rat CX3CL1/Fractalkine Chemokine Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF537) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Rat CX3CL1/Fractalkine Chemokine Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF537) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Rat CX3CL1/Fractalkine Chemokine Domain Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse brain (cingulate cortex)

Western Blot

0.1 µg/mL
Sample: Recombinant Rat CX3CL1/Fractalkine (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.

Rat CX3CL1/Fractalkine Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Rat CX3CL1/Fractalkine Chemokine Domain Biotinylated Antibody (Catalog # BAF537)
  • Standard: Recombinant Rat CX3CL1/Fractalkine Protein (Catalog # 536-FR)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4 using AF537 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

Alternate Names

FKN, Fractalkine, Neurotactin

Entrez Gene IDs

6376 (Human); 20312 (Mouse); 89808 (Rat); 102117496 (Cynomolgus Monkey)

Gene Symbol

CX3CL1

UniProt

O55145

Additional CX3CL1/Fractalkine Products

Product Documents for Rat CX3CL1/Fractalkine Chemokine Domain Antibody

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Product Specific Notices for Rat CX3CL1/Fractalkine Chemokine Domain Antibody

For research use only

Citations for Rat CX3CL1/Fractalkine Chemokine Domain Antibody

Customer Reviews for Rat CX3CL1/Fractalkine Chemokine Domain Antibody (2)

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  • Rat CX3CL1/Fractalkine Chemokine Domain Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Cortical neurons
    Species: Rat
    Verified Customer | Posted 12/19/2016
    Results published in: Article title: Noradrenaline induces CX3CL1 production and release by neurons Article reference: NP6524 Journal title: Neuropharmacology Final version published online: 08-Dec-2016 DOI information: 10.1016/j.neuropharm.2016.12.001
    Rat CX3CL1/Fractalkine Chemokine Domain Antibody AF537
  • Rat CX3CL1/Fractalkine Chemokine Domain Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Brain (cortex) tissue
    Species: Rat
    Verified Customer | Posted 12/01/2016
    Traumatic brain injury site in rat brain Fractalkine antibody (red) and DAPI (blue)
    Rat CX3CL1/Fractalkine Chemokine Domain Antibody AF537

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