Application

Fibronectin solution from human fibroblasts is used for the following applications:It is used in purification of human extracellular matrix (ECM) Coating of tissue culture plates with extracellular matrix componentsUsed during cell culture (For xeno-free, feeder-free culture the cells were passaged in to human fibronectin along with other components)Fibronectin from human plasma can be used with epithelial cells, mesenchymal cells, neuronal cells, fibroblasts, neural crest cells, and endothelial cells. It is recommended for use as a cell culture substratum at 1-5 μg/cm² or 0.5-50 μg/mL. The optimal concentration does depend on cell type as well as the application and research objectives.

Biochem/physiol Actions

Fibronectin is a high molecular mass extracellular matrix glycoprotein involved in many cellular processes, including tissue repair, embryogenesis, blood clotting, cell migration/adhesion, and embryogenesis and development. It is used in vitro as a substrate to enhance adherence and proliferation of many cell types. It has the capacity to bind to macromolecules such as collagen, fibrin, glycosaminoglycans, and Clq, apart from some bacteria.

Caution

The solution should be stored at -20°C. Under these conditions, the product is stable for at least 2 years. Vortexing, excessive agitation, or additoinal freezing and thawing of reconstituted fibronectin are not recommended.

Components

Fibronectin is an adhesive glycoproteins found in both the cell surface and in plasma.

General description

The matrix protein fibronectin consists of three units that are named Fn1, Fn2 and Fn3. The plasma protein has a molar mass of 31kDa. It is present in loose connective tissue, basement membranes, and the surface of various cell types.

Fibronectin 1 is a glycoprotein of the extracellular matrix that is coded by FN1 gene. It is expressed in the plasma and at the cell surface. It is mapped to human chromosome 2q35.

Preparation Note

This product is produced by human fibroblasts and then purified biochemically. It is supplied as a sterile solution at 0.5 mg/mL in a CAPS saline buffer. Before use, the product should be thawed slowly at 2-8°C, forming a clear solution. Avoid any vortexing or vigorous shaking of the solution, which will cause any fibronectin to “crash” out of solution. To coat plates, dilute this product in sterile HBSS solutions and coat with a minimal volume. Then, incubate at 37°C for 1-2 hours and wash 3 times with sterile HBSS and plate cells.