Description
Principle of the assay: human Fractalkine ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Fractalkine has been precoated onto 96-well plates. Standards(NSO,Q25 - R339) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Fractalkine is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Fractalkine amount of sample captured in plate.
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-aminoacid 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 adhesiveand migratory functions by interacting with the chemokine receptor CX3CR1.3 Its gene is located on humanchromosome 16 along with some CC chemokines known as CCL17 and CCL22.2, 4 It can act as a mediator ofsmooth muscle cell migration in human atherosclerosis, rather than mediate inflammatory cell recruitment.5