Fibronectin is a 450 kDa glycoprotein involved in cell adhesion and migration processes including embryogenesis, metastasis, wound healing, blood coagulation and host immunity. Fibronectin may be used as a coating on tissue culture surfaces to promote attachment, spreading and proliferation of various cell types.
- Target Species
- Product Form
- Purified bovine fibronectin - sterile liquid
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- None present
- >/=95% by SDS PAGE
- Approx. Protein Concentrations
- Protein concentration 0.5 mg/ml
- Endotoxin Level
- <10 EU/mg
- This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the protein. Storage in frost-free freezers is not recommended.
- 12 months from date of despatch
- Entrez Gene
- GO Terms
cell-substrate junction assembly
regulation of cell shape
apical plasma membrane
peptidase activator activity
- For research purposes only
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Applications of Fibronectin
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Instructions For Use
- Preparation of Fibronectin:
Generally fibronectin should be stored frozen or refrigerated at 1-5 mg/ml. If frozen, thaw at 37oC. Fibronectin should never be vortexed or treated roughly as it precipitates out of solution easily (visible as a granulated precipitate or as a slimy strand or clumps). Any observed insoluble aggregates are very difficult to resuspend. If precipitation is observed, the solution should initially be warmed to 37oC with gentle shaking, followed by aseptic filtration via a 5 µm (or smaller) cut off syringe filter.
It is also recommended that fibronectin should not be stored long term in buffer solutions containing Mg2+ or Ca2+ as these can contribute to fibronectin precipitation over time.
General Coating Procedure (for 96-well plates):
1. Plates: Choice of plates can affect the amount of protein successfully coated. Bio-Rad recommends the use of commercially available high protein binding plates.
2. Add between 1-10 µg/ml of fibronectin in PBS buffer for an overnight (or longer) incubation at 2-8oC (100 µl/well). The optimum amount of protein to saturate the plate surface should be determined by the user. For ELISA applications some users have coated with carbonate buffer pH 9.0 but this is generally not necessary and can damage the cell binding activity of the fibronectin.
3. Regardless of assay type (cell binding, ELISA etc.), it is necessary to block the remaining protein adherence sites on the plate. Therefore, in a separate step add a blocking protein such as 2-5% BSA in PBS for at least 1 hr at room temperature or overnight at 2-8oC (200 µl/well). Caution: The BSA solution should be 0.2 µm filtered prior to use to remove excess non-specific binding effects in the assay caused by insoluble BSA clumps. The use of RIA-grade (radio-immunoassay grade, suitable for immunoassays) BSA is recommended.
4. Blocked plates can stored in the refrigerator for several weeks or can be decanted and dried and stored for months in a dessicator. Dessicated material should be rehydrated for 15 min with PBS before use.
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