| Source: |
Escherichia coli |
| Molecular Weight: |
Approximately 62.6 kDa, a single non-glycosylated polypeptide chain containing 574 amino acids. |
| Purity: |
> 95% by SDS-PAGE and HPLC analyses. |
| Biological Activity: |
Measured by its ability to support cell attachment and spreading when used as a substratum for cell culture. In this application, the recommended concentration for this effect is typically 1-5 μg/cm2. Fibronectin can also be added to the media to support cell spreading at a concentration of 0.5-50 μg/mL. |
| Physical Appearance: |
Sterile filtered white lyophilized (freeze-dried) powder. |
| Formulation: |
Lyophilized from a 0.2 µm filtered concentrated solution in 20 mM Tris-HCl, 150 mM NaCl, 5% Trehalose, 0.02% Tween-20, pH8.5. |
| Endotoxin: |
Less than 0.1 EU/µg of rPrimeNectin GMP as determined by LAL method. |
| Reconstitution: |
Prior to opening, it is recommended to centrifuge the vial briefly to bring the contents down the bottom. Reconstitute in sterile PBS to a concentration of 0.1-5.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤ -20°C. Further dilutions should be made in appropriate buffered solutions. |
| Stability & Storage: |
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
A minimum of 12 months from date of receipt, when stored at ≤ -20°C as supplied.
1 month, 2 to 8°C under sterile conditions after reconstitution.
3 months, -20 to -70°C under sterile conditions after reconstitution.
Refer to lot-specific CoA for the Expiry Date. |
| Background: |
PrimeNectin, a derivative of recombinant human fibronectin, is a modified fragment of this large modular glycoprotein that serves as a ligand for various molecules, including fibrin, heparin, chondroitin sulfate, collagen/gelatin, and integrins. It plays a pivotal role in numerous cellular processes such as cell adhesion/migration, blood clotting, morphogenesis, tissue repair, and cell signaling. Fibronectin comprises several functional domains, notably the cell-binding domain (C-domain), heparin-binding virus adhesion domain (H-domain), and CS-1 sequence.
PrimeNectin facilitates retroviral-mediated gene transduction by promoting the co-localization of target cells and virions. Specifically, the viral particle can adhere to the virus adhesion region (H-domain) of fibronectin, while target cells primarily bind via the interaction of cell surface integrin receptors VLA-5 and VLA-4 with the fibronectin C-domain and CS-1 site, respectively. This proximity between cells and viruses enhances the efficiency of virus infection. When PrimeNectin is applied as a coating on the surface of a cell culture dish, it significantly boosts the transfection capacity of both mammalian and insect cells. |
