Analysis Note

The biological activity of recombinant human fibronectin was tested in culture by measuring the ability of immobilized fibronectin to support adhesion of CHO cells.

Application

Fibronectin human has been used for coating cell culture plates for culturing endothelial progenitor cells. It has also been used as a component of Dulbecco′s modified Eagle′s medium (DMEM) in the in vitro matrigel invasion assay of glioblastoma cells.

Epithelial cells, mesenchymal cells, neuronal cells, fibroblasts, neural crest cells, endothelial cells. Recommended for use as a cell culture substratum at 1-5 µg/cm² or 0.5-50 µg/ml. Optimal concentration depends on cell type as well as the application or research objectives.

Biochem/physiol Actions

Fibronectin (FN) is essential for embryonic development and mediates cell adhesion and migration functions. It mediates apoptosis in nasopharyngeal carcinoma (NPC). The plasma FN regulates thrombosis. FN acts as a ligand for the integrin receptor family and contains collagen, glycoprotein and glycosaminoglycan binding sites. Mutation in the FNI gene leads to an imbalance in the soluble and insoluble forms of fibronectin leading to glomerular deposits in hereditary kidney disease, glomerulopathy. Missense mutations in the FN1 gene are implicated in the skeletal dysplasias disorder, Spondylometaphyseal dysplasias.

General description

Fibronectin (FN) is a glycoprotein with 5-9% carbohydrates present as N- and O-linked oligosaccharides. It belongs to the ligand glycoprotein family. The FN gene is localized to human chromosome 2q35. FN exists as two major isoforms, plasma-based soluble and extracellular matrix (ECM)-based insoluble form. FN is a multidomain protein and comprises repeats of FNI, FNII and FNIII repeats. It also has fibrin-binding sites and exists as a dimer. Alternative splicing of the FN1 gene leads to the generation of many isoforms. Recombinant human fibronectin is expressed in human 293 cells as a glycoprotein with a calculated molecular mass of 259.5 kDa. This protein is manufactured in human cells using an all-human production system with no serum. The human cells expression system allows human-like glycosylation and folding, and often supports better stability of the protein in culture.

Physical form

Supplied as a powder, lyophilized from CAPS buffered saline.