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Fibronectin EIA Kit



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Product Name: Fibronectin EIA Kit

# of Samples: 1X96 Assays

Intended Use: The human Fibronectin EIA Kit is an in vitro enzyme immunoassay (EIA) kit for the specific quantitative determination of human Fibronectin (hFN) in serum, urine, cell culture supernatants, and other biological fluids. This kit is suitable for quantitation of soluble human Fibronectin.
This kit is for research use only. It is not for use in diagnostic or therapeutic procedures.

Introduction: The adhesive glycoprotein fibronectin (FN), Mr440,000, is widely distributed on cell surfaces, in the extracellular matrix, and in plasma. FN is involved in a variety of cellular processes, including cell-to-substrate adhesion, cell migration, and regulation of cell morphology (1). Webb et al first reported in 1980 (2) that the amount of fragmented FN in the urine of patients with prostatic cancer was increased compared to normal subjects. Another group has reported that an FN-like protein, with a low molecular mass than plasma FN, was increased in sera from patients with malignant disease (3). In particular, many examinations have been done to evaluate the clinical usefulness of measurements of plasma FN (4-7). Recently, Katayama et al reported that urinary FN (UFN) in cancer patients was almost always present as fragments of different sizes containing the central “cell-binding domain”, and that a polyclonal antibody raised against native plasma FN could not react well with these fragmented UFN. They demonstrated that two monoclonal antibodies, reactive with the “cell-binding domain” (8) are suitable for in vitro assays of FN in biological fluids and FN-fragments excreted in the urine of cancer patients, and that significantly higher levels of UFN were observed in many kinds of cancer (9,10).

Principle of the test: The human Fibronectin EIA Kit is a solid phase EIA based on a sandwich method that utilizes two mouse monoclonal anti-human Fibronectin antibodies to detect Fibronectin by two-step procedure. One of the mouse monoclonal anti-human Ffibronectin antibodies is bound to a microtiter plate to create the solid phase. Nonspecific binding is blocked by a blocking buffer. Samples and standards are incubated in microtiter-plate wells. The second step is to wash the plate and add a second anti- Fibronectin antibody labelled with peroxidase (POD). During the incubation, human Fibronectin, bound to anti-human Fibronectin antibody (solid phase) on one side, is tagged on the other by POD-anti-human Fibronectin antibody. The reaction between POD and substrate (H2O2 and tetramethylbenzidine) results in colour development with intensities proportional to the amount of human Fibronectin present in samples and standards. The amount of human Fibronectin can be quantitated by measuring the absorbance using an EIA plate reader. Accurate sample concentrations of human Fibronectin can be determined by comparing their specific absorbances with those obtained for the standards plotted on a standard curve.

Storage and Stability: This kit is shipped at 2 - 8C and should be stored at 2 - 8C if not used. Under this condition, the kit is stable until the expiration date on label.

Sensitivity and Specificity: This kit specifically measures human Fibronectin. This kit cannot be used to measure mouse, bovine, rabbit, porcine Fibronectin. The application of this kit for quantitating Fibronectin from other sources has not been tested.

Product Name: Fibronectin EIA Kit

Fibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin. Fibronectins are involved in cell adhesion, cell motility, opsonization, wound healing, and maintenance of cell shape. Interaction with TNR mediates inhibition of cell adhesion and neurite outgrowth (By similarity).

Subunit: Mostly heterodimers or multimers of alternatively spliced variants, connected by 2 disulfide bonds near the carboxyl ends; to a lesser extent homodimers. Interacts with FBLN1, AMBP, TNR, LGALS3BP and COL13A1. Interacts with FBLN7 (By similarity).

Subcellular Location: Secreted, extracellular space, extracellular matrix.

Tissue Specificity: Plasma FN (soluble dimeric form) is secreted by hepatocytes. Cellular FN (dimeric or cross-linked multimeric forms), made by fibroblasts, epithelial and other cell types, is deposited as fibrils in the extracellular matrix. Ugl-Y1, Ugl-Y2 and Ugl-Y3 are found in urine.

Developmental Stage: Ugl-Y1, Ugl-Y2 and Ugl-Y3 are present in the urine from 0 to 17 years of age.

Ptm: Sulfated.

Ptm: It is not known whether both or only one of Thr-2064 and Thr-2065 are/is glycosylated.

Ptm: Forms covalent cross-links mediated by a transglutaminase, such as F13A or TGM2, between a glutamine and the epsilon-amino group of a lysine residue, forming homopolymers and heteropolymers (e.g. fibrinogen-fibronectin, collagen-fibronectin heteropolymers).

Disease: Defects in FN1 are the cause of glomerulopathy with fibronectin deposits type 2 (GFND2) [MIM:601894]; also known as familial glomerular nephritis with fibronectin deposits or fibronectin glomerulopathy. GFND is a genetically heterogeneous autosomal dominant disorder characterized clinically by proteinuria, microscopic hematuria, and hypertension that leads to end-stage renal failure in the second to fifth decade of life.

Similarity: Contains 12 fibronectin type-I domains.

Similarity: Contains 2 fibronectin type-II domains.

Similarity: Contains 16 fibronectin type-III domains.

Additional Information

Name Fibronectin EIA Kit
Related Product Names Fibronectin EIA ELISA Kit; FN; Cold-insoluble globulin; CIG FNFibronectin EIA KitFN1
NCBI Acc Number NP_002017.1
Molecular Weight 262607
Datasheets / Downloads GWB-F363CE Datasheet
Swiss Prot Number P02751
Intended Use Research Use Only