Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Bovine, Hamster, Monkey

Applications

Validated:

Western Blot, Neutralization

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Bioassay, ELISA Capture, ELISA Development

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CX3CL1/Fractalkine Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human CX3CL1/Fractalkine
Gln25-Gly100
Accession # Q6I9S9

Specificity

Detects human CX3CL1/Fractalkine in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant human (rh) MCP‑2 and rhMCP-4 is observed.

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 Human CX3CL1/Fractalkine Chemokine Domain Antibody

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

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

Recombinant Human CX3CL1/Fractalkine (Catalog # 365-FR) chemoattracts the BaF3 mouse pro-B cell line transfected with mouse 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 Human CX3CL1/Fractalkine (100 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CX3CL1/Fractalkine Chemokine Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF365). The ND50 is typically 0.75-4.5 µg/mL.
Detection of CX3CL1/Fractalkine by Western Blot

Detection of CX3CL1/Fractalkine by Western Blot

CX3CR1 is required for keratinocytes differentiation. (A) Expression of CX3CR1 in parental PA-1 cell line&that stably transfected with either CX3CR1 or scrambled shRNA constructs was examined with WB. Actin served as a loading control. CX3CR1 expression levels quantified using digital densitometry&normalized to the levels of actin expression. (B) Expression of cytokeratin 14 in PA-1 cells stably transfected with either CX3CR1 or scrambled shRNA constructs treated with recombinant BMP-4 or vehicle, as indicated, was examined with immunocytochemistry. CK14 in PA-1 was probed with anti-CK14&Alexa555-conjugated anti-rabbit antibodies,&DNA was stained with 4’,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI); CK14 – red, DNA – blue. Images taken using 5× magnification on the objective. The histogram demonstrates the average intensity of CK14 signal across the field as determined by the Zeiss AxioVision software. Data is an average of three independent experiments. *p < 0.05. (C) Expression of cytokeratins 14&18 in PA-1 cells stably transfected with either CX3CR1 or scrambled shRNA constructs treated with recombinant BMP-4 or vehicle, as indicated, was examined with WB. beta -Tubulin served as a loading control. The histogram shows CK14&CK18 expression levels. Expression of CK18 in CX3CR1shRNA-transfected PA-1 cells was arbitrarily set as 1&expression of both CK14&CK18 in other conditions was calculated accordingly. The data represent a typical WB image,&quantitative analysis was performed using three independently performed experiments. *p < 0.05. (D) Expression of CX3CL1 in SKOV-3&PA-1 cell lines was examined with WB. Actin served as a loading control. SKOV-3 cell line was used as a positive control. Image collected&cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23958497), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human CX3CL1/Fractalkine Chemokine Domain Antibody

Application
Recommended Usage

Western Blot

0.1 µg/mL
Sample: Recombinant Human CX3CL1/Fractalkine Chemokine Domain (Catalog # 362-CX)

Neutralization

Measured by its ability to neutralize CX3CL1/Fractalkine-induced chemotaxis in the BaF3 mouse pro‑B cell line transfected with mouse CX3CR1. The Neutralization Dose (ND50) is typically 0.75-4.5 µg/mL in the presence of 100 ng/mL Recombinant Human CX3CL1/Fractalkine.

Reviewed Applications

Read 3 reviews rated 4.3 using AF365 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 novel chemokine identified through bioinformatics. CX3CL1 has a unique C-X3-C cysteine motif near the amino-terminus and is the first member of a fourth branch of the chemokine superfamily. Unlike other known chemokines, CX3CL1 is a type 1 membrane protein containing a chemokine domain tethered on a long mucin-like stalk. Human CX3CL1 cDNA encodes a 397 amino acid (aa) residue membrane protein with a 24 aa residue predicted signal peptide, a 76 aa residue chemokine domain, a 241 aa residue stalk region containing 17 degenerate mucin-like repeats, a 19 aa residue transmembrane segment and a 37 aa residue cytoplasmic domain. The extracellular domain of human CX3CL1 can be released, possibly by proteolysis at the dibasic cleavage site proximal to the membrane, to generate soluble CX3CL1. CX3CL1 mRNA has been detected in various tissues including the brain and heart. The expression of CX3CL1 was also reported to be up-regulated in endothelial cells and microglia by inflammatory signals. 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. The gene for human CX3CL1 has been mapped to chromosome 16q.

References

  1. Pan, Y. et al. (1997) Nature 387:611.
  2. Bazan, J.F. et al. (1997) Nature 385:640.
  3. Mackay, C.R. (1997) Current Biology 7:R384.

Alternate Names

FKN, Fractalkine, Neurotactin

Entrez Gene IDs

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

Gene Symbol

CX3CL1

UniProt

Q6I9S9

Additional CX3CL1/Fractalkine Products

Product Documents for Human CX3CL1/Fractalkine Chemokine Domain Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CX3CL1/Fractalkine Chemokine Domain Antibody

For research use only

Citations for Human CX3CL1/Fractalkine Chemokine Domain Antibody

Customer Reviews for Human CX3CL1/Fractalkine Chemokine Domain Antibody (3)

4.3 out of 5
3 Customer Ratings
5 Stars
33%
4 Stars
67%
3 Stars
0%
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1 Stars
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Customer Images

Showing  1 - 3 的 3 reviews Showing All
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  • Human CX3CL1/Fractalkine Chemokine Domain Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Pancreas tissue
    Species: Human
    Verified Customer | Posted 09/13/2023
    FFPE antigen retrieval in pH 9 buffer primary antibody x100 for 1h biotinylated rabbit anti-goat and streptavidin-HRP
    Human CX3CL1/Fractalkine Chemokine Domain Antibody AF365
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 22943205
    Species: Human
    Verified Customer | Posted 01/08/2015
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 23470165
    Species: Human
    Verified Customer | Posted 01/08/2015

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