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EK000023-EK0349: Human Fibronectin ELISA Kit
Range 156pg/ml-10ng/ml
Sensitivity < 10pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of human Fibronectin in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Fibronectin (FN) also known as LETS, is identified on the surfFN of fibroblasts by labeling with radioactive compounds or specific antibodies. Fibronectin is a 430,000-dalton dimeric glycoprotein that exists in 2 forms, termed cellular and plasma fibronectin. Cellular and plasma fibronectins are heterodimers consisting of similar but not identical polypeptides.1 These two forms of FN differ in biologic activity. Fibronectins bind cell surfFNs and various compounds including collagen, fibrin, heparin, DNA, and actin. Because fibronectin stimulates endocytosis in several systems and promotes the clearance of particulate material from the circulation, it could function in the clearance of C1q-coated material such as immune complexes or cellular debris.2 Fibronectins are involved in cell adhesion, cell motility, opsonization, would healing, and maintenance of cell shape. LETS, encoded on chromosome 8, is responsible for the LETS protein expression in humans. Because LETS has been implicated in tumorigenicity and cellular transformation, it is of interest that rearrangement or modifications in the number of chromosome 8 have been associated with certain forms of cancer.3 The standard product used in this kit is isolated from rat plasma with the molecular mass of 200-250KDa.
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. Kornblihtt, A. R.; Umezawa, K.; Vibe-Pedersen, K.; Baralle, F. E. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J. 4: 1755-1759, 1985.
2. Bing, D. H.; Almeda, S.; Isliker, H.; Lahav, J.; Hynes, R. O. Fibronectin binds to the C1q component of complement. Proc. Nat. Acad. Sci. 79: 4198-4201, 1982.
3. Owerbach, D.; Doyle, D.; Shows, T. B. Genetics of the large, external, transformation-sensitive (LETS) protein: assignment of a gene coding for expression of LETS to human chromosome 8. Proc. Nat. Acad. Sci. 75: 5640-5644, 1978.
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