Natural and recombinant human Fractalkine. This assay detects all forms of human Fractalkine containing the chemokine domain.
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Human Fractalkine immunoassay is a 4.5 hour solid phase ELISA designed to measure Fractalkine in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains NS0-expressed recombinant human Fractalkine and has been shown to accurately quantitate the recombinant factor. Results obtained using natural Fractalkine showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Fractalkine.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Serum, EDTA Plasma, Heparin Plasma, Urine, Human Milk
Cell Culture Supernates, Saliva
The recovery of Fractalkine spiked to three different levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples containing or spiked with high concentrations of Fractalkine were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
CX3CL1/Fractalkine is a transmembrane adhesion protein with a chemokine domain separated from the membrane by a mucin stalk. It is upregulated on many cell types during inflammation. The chemokine and mucin regions can be shed as a soluble chemokine that signals through the receptor CX3CR1. During extravasation, membrane-bound CX3CL1 traps leukocytes, then is cleaved to allow diapedesis. Soluble CX3CL1 protects neurons from microglial neurotoxicity, recruits macrophages for wound healing, recruits osteoclast precursors for bone resorption, and contributes to pathogenesis in coronary artery disease.
Entrez Gene IDs
6376 (Human); 20312 (Mouse); 89808 (Rat);
ABCD-3; C3Xkine; chemokine (C-X3-C motif) ligand 1; CX3C membrane-anchored chemokine; 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;
Refer to the product for complete assay procedure.
The conjugate must be kept cold during use. Bring all other reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample
Add 100 µL of Standard, control, or sample to each well within 15 minutes. Cover with a plate sealer, and incubate at 2-8 °C for 3 hours.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL cold Conjugate
Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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