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Rat CX3CL1/Fractalkine Chemokine Domain Antibody

R&D Systems, part of Bio-Techne | Catalog # AF537

R&D Systems, part of Bio-Techne
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AF537
AF537-SP

Key Product Details

Species Reactivity

Validated:

Rat

Cited:

Mouse, Rat

Applications

Validated:

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

Cited:

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

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

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.

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.

Reconstitution Buffer Available:
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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.

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

Additional CX3CL1/Fractalkine Products

Product Documents for Rat CX3CL1/Fractalkine Chemokine Domain Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Rat CX3CL1/Fractalkine Chemokine Domain Antibody

For research use only

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