Cellular Fibronectin ELISA Kit (cFN)

Full Name: Cellular Fibronectin ELISA Kit (cFN)
Reactivity: Human
Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
Sensitivity: 0.469 ng/ml

INTRODUCTION

Cellular fibronectin, often referred to as cFN is a glycoprotein, with a molecular weight that exists in the extracellular matrix. In all vertebrates it acts as a regulator of structure and adhesion.

Different types of cells secrete fibronectin which then forms fibrils outside the cells. These fibrils bind to integrins on the surface of cells and other components within the matrix like collagen, fibrin and heparan sulfate proteoglycans. This intricate network of fibronectin fibrils provides a scaffold in the environment that helps organize cells and promote attachment. During development fibronectin is crucial for functioning as defects can lead to embryonic lethality.

The structure of cFN consists of domains that play different roles including binding to receptors on cell surfaces or other proteins within the extracellular matrix and even interacting with other regions within fibronectin itself. Structurally speaking, cFN is composed of two subunits weighing around 250 kDa each that are closely identical to each other. These subunits are linked together by disulfide bonds. Each subunit contains repeating modules known as types I, II and III which have sites interacting with integrins found on cell surfaces like α5β1. One notable integrin binding domain is the amino acid sequence RGD (arginine acid). Overall, the modular nature of cFN allows it to interact with ligands, due to its structure.

INTENDED USE

Human cellular fibronectin ELISA kit can measure concentrations of cFN (cellular fibronectin) present is plasma, serum, biological fluids or tissue homogenate samples.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti-cFN antibody.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this cellular fibronectin ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 1.562, 3.125, 6.25, 12.5, 25, 50, 100 ng/ml.
Reactivity: Human
Sensitivity: 0.938 ng/ml
Range: 1.563 – 100 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for cFN, no cross reactivity or interference between cFN and analogues was detected.
– Recovery: Serum (85 – 102%), EDTA Plasma (87 – 103%), Heparin Plasma (86 – 103%).
– Linearity: Serum (82 – 100%), EDTA Plasma (86 – 104%), Heparin Plasma (95 – 104%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. Interplay between elevated cellular fibronectin and plasma fibrin clot properties in type 2 diabetes. Thromb Haemost. (2017) 117 (9): 1671-1678. Konieczyńska M., et al.
2. Fibronectin maintains the balance between hemostasis and thrombosis. Cell Mol Life Sci. (2016) 73 (17): 3265-77. Wang Y. and Ni H.
3. Differences in domain structure between human fibronectins isolated from plasma and from culture supernatants of normal and transformed fibroblasts. Studies with domain-specific antibodies. J Biol Chem. (1985) 260 (8): 5105-14. Sekiguchi K., et al.
4. Cellular fibronectin in serum and plasma: a potential new tumour marker? Br J Cancer. (1995) 71 (3): 578-82. Ylätupa S., et al.

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