Fibronectin (FN) is an extracellular glycoprotein that exists in soluble form in body fluids or in insoluble form in the extracellular matrix. As one of the main cell adhesion molecules, FN plays a key role in many important physiological processes, such as embryogenesis, wound healing, hemostasis and thrombosis. Changes in fibronectin expression, degradation and combination are closely related to a large number of pathological occurrences, including cancer and fibrosis. Applications of fibronectin (FN): 1) As a cell culture matrix; its function is to promote cell growth, increase cell adhesion rate, enhance cell metabolism level, shorten cell growth time; increase cell fusion rate in hybridoma technology, etc. 2) Disease diagnosis and treatment; its function is wound repair and healing, cancer diagnosis and treatment, treatment of vascular system and cardiovascular and cerebrovascular diseases, etc. This product is a recombinant human fibronectin derived from rice endosperm cells, existing in monomeric form, and can be used as a cell culture matrix to replace Matrigel, rat tail collagen, blood-derived fibronectin, etc., to increase cell adhesion rate, enhance cell metabolism level, and shorten cell growth cycle. It can also be added directly to the culture medium to replace or reduce the amount of serum used, promote cell adhesion, migration, mobile growth, and maintain cell status. This product is a freeze-dried powder, which is of cell culture grade and has the characteristics of small batch-to-batch differences and no potential risk of viral infection. Product Properties CAS Number 86088-83-7 Source Rice Molecular Weight 200~210 kDa Physical Appearance Sterile Filtered White lyophilized (freeze-dried) powder. Purity > 95% by SDS-PAGE pH 6.0~8.0 Biological Activity The ED50 as determined by a celI adhesion assay using B16-F1 mouse melanoma cells is less than 1.1 μg/mL,corresponding to a specific activity of > 1.17 X 107 IU/mg. Endotoxin < 0.5 EU/μg Shipping and Storage This product should be stored at -25~-15℃ for 3 years. Figures Figure 1. The HPLC purity of fibronectin is more than 95%. Figure 2. Activity analysis of fibronectin on CHO-K1 cell proliferation Figure 3. CHO-K1 cells growth was significantly promoted by fibronectin protein. Reconstitution Dissolve recombinant human fibronectin (rFN) in 1×PBS to prepare a storage solution with a concentration of 1 mg/mL. Store at -20°C in aliquots according to single-use doses to avoid repeated freezing and thawing. It is effective for 3-6 months. Instructions for use 1. Coating experiment (taking 6-well plate as an example, for other culture plates, please refer to Table 1 ) 1.1 Take out an appropriate amount of recombinant human fibronectin (rFN) and dilute it to the working solution concentration with sterile 1×PBS, pH 7.2~7.4, and the final concentration reference range is 50~100 μg/mL. After mixing, add it to the 6-well plate, 800 μL/well, to ensure that the working solution completely covers the bottom of the 6-well plate. 1.2 Place the 6-well plate with rFN working solution at 4 ℃ overnight or 37 ℃ for coating for 1 h. 1.3 Collect the working solution in the 6-well plate and store it at 4 ℃. It can be used for coating other culture bottles/plates within one week. 1.4 Take out the 6-well plate coated with rFN, add normal cell culture medium, and directly inoculate cells. rFN promotes cell adhesion and growth. 1.5 If the coated 6-well plate is not used for the time being, store in sealed container at 4℃. It is recommended to use it within one week. Table 1. Recommended rFN addition amounts for different culture bottles/plates Cell culture dishes/plates Bottom area (cm2) Recommended dosage 96-well plate 0.32 50μL/well 24-well plate 2 200μL/well 12-well plate 4.5 400μL/well 6-well plate 9.6 800μL/well 35mm dish 8 600μL/well 60mm dish 21 1.6mL/dish T-25 bottle 25-28 2 mL/ bottle In the coating/plating experiment, the amount of recombinant human fibronectin (rFN) working solution added should be adjusted according to the size of the culture dish/bottle, as shown in Table 1 below. 2. Cell Culture Experiment During cell culture, adding rFN directly to the cell culture medium can promote cell adhesion, migration, and maintain cell status. The final concentration reference range is 5~50 μg/mL. Notes 1. This product is for research use only. 2. Please operate with lab coats and disposable gloves，for your safety. Cautions 1. Avoid repeated freeze-thaw cycles. 2. For your safety and health, please wear lab coats and disposable gloves for operation. 3. For research use only! Citations & References: [1] Qin, W., Tian, Y., Zhang, J. et al. The double inhibition of PDK1 and STAT3-Y705 prevents liver metastasis in colorectal cancer. Sci Rep 9, 12973 (2019). [2] Zhang, Wenhao et al. Inhibition of Apoptosis Reduces Diploidization of Haploid Mouse Embryonic Stem Cells during Differentiation. Stem Cell Reports, Volume 15, Issue 1, 185 - 197 [3] Wang, Yuan et al. Anti photoaging mechanism of a novel recombinant human fibronectin peptide (rhFNP) derived from the extracellular matrix. Heliyon, Volume 11, Issue 4, e42730 FAQ Q: Does this product come with a label? A: This product does not come with a label. Q: Can this product be dissolved using PBS buffer solution? A: You can dissolve it using PBS with a pH of 7.2. Documents Manuals
