Fibronectin EIA Kit

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

Fibronectin EIA Kit

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).

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.

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

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.

Contains 2 fibronectin type-II domains.