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EK000083-EK0357: Mouse Fractalkine/CX3CL1 ELISA Kit
Range 156pg/ml-10,000pg/ml
Sensitivity < 10pg/ml
Specificity: No detectable cross-reactivity with Eotaxin, MCP-1, MCP-2, 6Ckine, MARC.
Application: For quantitative detection of mouse Fractalkine in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Chemokine (C-X3-C motif) ligand 1 (CX3CL1) is a large cytokine protein of 373 amino acids, it contains multiple domains and is the only known member of the CX3C chemokine family. It is also commonly known under the names fractalkine (in humans) and neurotactin (in mice).1, 2 CX3CL1 is produced as a long protein (with 373-amino acid in humans) with an extended mucin-like stalk and a chemokine domain on top. The mucin-like stalk permits it to bind to the surface of certain cells. However a soluble (90 kD) version of this chemokine has also been observed. Soluble CX3CL1 potently chemoattracts T cells and monocytes, while the cell-bound chemokine promotes strong adhesion of leukocytes to activated endothelial cells, where it is primarily expressed.2 CX3CL1 elicits its adhesive and migratory functions by interacting with the chemokine receptor CX3CR1.3 Its gene is located on human chromosome 16 along with some CC chemokines known as CCL17 and CCL22.2, 4 It can act as a mediator of smooth muscle cell migration in human atherosclerosis, rather than mediate inflammatory cell recruitment.5 The standard product used in this kit is recombinant human CX3CL1, consisting of 76 amino acids with the molecular mass of 8.5kDa. and mouse CX3CL1, consisting of 84 amino acids with the molecular mass of 9.5kDa.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Pan et al. Neurotactin, a membrane-anchored chemokine upregulated in brain inflammation. Nature 387: 611-617, 1997.
2. Bazan et al. A new class of membrane-bound chemokine with a CX3C motif. Nature 385: 640-644, 1997.
3. Imai et al. Identification and molecular characterization of fractalkine receptor CX3CR1, which mediates both leukocyte migration and adhesion. Cell 91: 521-530, 1997.
4. Nomiyama et al. Human chemokines fractalkine (SCYD1), MDC (SCYA22) and TARC (SCYA17) are clustered on chromosome 16q13. Cytogenet. Cell Genet. 81: 10-11, 1998.
5. Lucas, A. D.; Bursill, C.; Guzik, T. J.; Sadowski, J.; Channon, K. M.; Greaves, D. R. : Smooth muscle cells in human atherosclerotic plaques express the fractalkine receptor CX3CR1 and undergo chemotaxis to the CX3C chemokine fractalkine (CX3CL1). Circulation 108: 2498-2504, 2003.
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