Fibronectin Recombinant
Escherichia Coli.
Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
Description
Fibronectin Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 574 amino acids and having a molecular mass of 62.6kDa. The Fibronectin is purified by proprietary chromatographic techniques.
Product Specs
Fibronectin is a glycoprotein that plays a crucial role in various cellular processes, including wound healing, embryonic development, blood coagulation, and cell migration/adhesion. Elevated levels of fibronectin in plasma are observed in individuals with severe coronary artery disease. Additionally, increased plasma fibronectin levels are associated with venous thromboembolism (VTE), particularly in men. This suggests a potential link between biomarkers, risk factors for arterial atherothrombosis, and VTE. Fibronectin exists in two primary forms: an insoluble glycoprotein dimer that acts as a connector in the extracellular matrix (ECM), and a soluble disulfide-linked dimer found in plasma. Hepatocytes produce the plasma form, while fibroblasts, chondrocytes, endothelial cells, macrophages, and certain epithelial cells synthesize the ECM form. Fibronectin also functions as a general cell adhesion molecule, facilitating cell attachment to collagen or proteoglycan substrates. By binding to various components of the ECM and membrane-bound fibronectin receptors on cell surfaces, fibronectin orchestrates cellular interaction with the ECM.
Recombinant Human Fibronectin, produced in E. coli, is a single, non-glycosylated polypeptide chain comprising 574 amino acids. It has a molecular weight of 62.6 kDa. The purification of recombinant human fibronectin is achieved using proprietary chromatographic techniques.
The product is lyophilized from a 0.2 µm filtered concentrated solution. The solution contains 20 mM Tris-HCl (pH 8.0), 150 mM NaCl, 5% Trehalose, and 0.02% Tween-20.
To reconstitute the lyophilized Fibronectin, it is recommended to dissolve it in sterile 18 MΩ-cm H₂O at a concentration of at least 100 µg/ml. This solution can then be further diluted into other aqueous solutions as needed.
The purity of the Fibronectin is determined using the following methods:
- Analysis by RP-HPLC
- Analysis by SDS-PAGE
The purity is determined to be greater than 95.0%.
Lyophilized Fibronectin remains stable at room temperature for a period of 3 weeks. However, for long-term storage, it is recommended to store the desiccated product at a temperature below -18°C. After reconstitution, Fibronectin should be stored at 4°C for a period of 2-7 days. For future use, store it below -18°C. It is important to avoid repeated freeze-thaw cycles.
The biological activity of the protein was assessed based on its capacity to facilitate cell attachment and spreading when employed as a substrate for cell culture. In this context, a concentration range of 1-5 µg/cm2 is generally recommended for achieving this effect. Alternatively, Fibronectin can be introduced to the media at a concentration of 0.5-50 µg/ml to support cell spreading. Determining the optimal concentrations for specific user applications will require individual optimization.
Escherichia Coli.
Product Science Overview
Fibronectin exists in two main forms:
- Soluble plasma fibronectin: Circulates in the blood and other body fluids.
- Insoluble cellular fibronectin: Forms a fibrillar network in the extracellular matrix.
Fibronectin is composed of two nearly identical polypeptide chains linked by disulfide bonds. Each chain contains multiple domains responsible for binding to other fibronectin molecules, cell surface receptors, and other extracellular matrix components .
Fibronectin is involved in several key biological processes:
- Cell Adhesion and Migration: Fibronectin binds to cell surface receptors called integrins, facilitating cell attachment and movement.
- Wound Healing: It plays a critical role in tissue repair by promoting cell migration to wound sites.
- Blood Clotting: Fibronectin interacts with fibrin during blood clot formation, aiding in the stabilization of the clot.
- Morphogenesis: It is essential for the development of tissues and organs during embryogenesis .
Recombinant human fibronectin is widely used in research and clinical settings:
- Cell Culture: It is used as a coating for cell culture dishes to promote cell attachment and spreading.
- Tissue Engineering: Fibronectin is utilized in the development of biomaterials for tissue regeneration.
- Drug Delivery: It can be incorporated into drug delivery systems to enhance the targeting and efficacy of therapeutic agents.
- Diagnostics: Fibronectin-based assays are used for the detection of various diseases .
The production of recombinant human fibronectin involves several steps:
- Gene Cloning: The gene encoding fibronectin is cloned into an expression vector.
- Expression: The vector is introduced into host cells, which produce the fibronectin protein.
- Purification: The protein is purified using techniques such as affinity chromatography to achieve high purity levels .
Recombinant human fibronectin is available in different formulations, including carrier-free versions that do not contain bovine serum albumin (BSA). This is particularly useful for applications where the presence of BSA could interfere with experimental results .
