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F2006 Sigma

Fibronectin from human plasma

lyophilized powder, BioReagent, suitable for cell culture

Synonym: CIG, Cold insoluble globulin

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Components

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

Application

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.

Fibronectin from human plasma is used for the following applications:
• Used in cell seeding and neuronal differentiation (as one of the components in expansion medium for the re-suspension of CBSCs (Cord blood stem cells) and is also used in differentiation medium)
• Used in cell culture (the culture chamber was coated with fibronectin for the expression of HEK293 cells)
• Used in murine embroyonic stem cell line (CCE) culture and induction of endothelial cell differentiation (for the induction of endothelial cell differentiation, cultured cells were pre-coated with fibronectin)
• In-vitro fibronectin (FN) pulldown assays
• Protein specificity
• Immunofluorescence (used in staining of cell layers for membrane type-1 matrix metalloproteinase (MT1-MMP) for coating of slides)
• Extracellular matrix binding assay
• Used in cell migration study

Caution

The reconstituted solution should be stored in working aliquots at -20°C or lower. Vortexing, excessive agitation, or additoinal freezing and thawing of reconstituted fibronectin are not recommended.

Preparation Note

This product is lyophilized from 0.05 M Tris buffered saline at pH 7.5. It is soluble at 1 mg/mL in water, and should be allowed to dissolve for at least 30 minutes at 37°C. A small amount of undissolved material may remain, but it will not affect performance. The protein homogeneity is evaluated by immunoelectrophoresis, is endotoxin tested, and the source material was tested negative for HIV, hepatitis B and hepatitis C. In coating culture surfaces, fibronectin should be diluted in sterile balanced sterile salt solution and coated with minimal volume. Surface should then be air dried for 45 minutes at room temp and can be stored for 2-4 weeks at 2-8°C.

Biochem/physiol Actions

Fibronectin (FN) plays an essential role in multiple biological functions such as, angiogenesis, cell migration and differentiation and wound healing. Studies in Chinese Han population show that mutations in this gene might be linked with susceptibility to knee osteoarthritis (OA). Splice variant extra domain-B (EDB) of FN is a hallmark of tumor angiogenesis, and with increased matrix stiffness there is an increase in the production of this isoform by endothelial cells. In colorectal cancer (CRC), EDA is linked with metastasis, vasculogenesis and tumorigenesis and promotes epithelial-mesenchymal transition.

General description

Fibronectin (FN) is a multifunctional glycoprotein, the gene of which is localized to human chromosome 2q34-36. This gene spans 75kb and contains 50 exons. It exists as two major isoforms, one soluble and one insoluble form. The former is present in plasma, whereas the insoluble form resides in tissues and extracellular matrix (ECM) of cartilage. There are around 20 different forms of FN due to alternative splicing of a variable region and two type III exons called Extra Domains A and B.

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Protocols & Articles

Articles

Fibronectin Cell Attachment Protocol

Freezing and thawing of reconstituted fibronectin is not recommended as breakdown of protein will occur.
George Sitterley
BioFiles 2008, 3.8, 9.
Keywords: Phase transitions, Tools

Fibronectin

Fibronectin (FN) is a multifunctional, extracellular matrix glycoprotein composed of two nearly identical disulfidebound polypeptides of molecular weight 220 kDa. Cellular fibronectin is structurally...
George Sitterley
BioFiles 2008, 3.8, 8.
Keywords: Adhesion, Cell culture, transformation

Plasma Derived Proteins and Enzymes

Despite their complexity, blood and plasma are abundant biological resources for the discovery of drug targets and biomarkers for human disease. It is estimated that plasma may contain as many as 40,...
BioFiles 2006, 1.5, 2.
Keywords: Applications, Gas chromatography, Type

The Cancer Stem Cell Hypothesis

Traditionally, cancer has been viewed as a disease in which environmental or endogenous events induce mutations to critical oncogenes and tumor suppressor genes within a normal cell. The clinical man...
Vicki Caligur
BioFiles 2008, 3.5, 4.
Keywords: Angiogenesis, Apoptosis, Asymmetric synthesis, Cancer, Catalysis, Cell culture, Cell division, Cell proliferation, Clinical, DNA replication, Degradations, Eliminations, Environmental, Events, Gastrointestinal, Gene expression, Growth factors, Ligands, Metabolic Pathways, Phosphorylations, Poisons, Transcription, Transfection, transformation

Peer-Reviewed Papers
15

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