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Active Fibronectin (FN)

FN1; CIG; FINC; LETS; MSF; GFND2; Anastellin; Migration-Stimulating Factor; Cold-Insoluble Globulin; Large, External, Transformation-Sensitive Protein

ACTIVITY TEST

Fibronectin (FN) is a high-molecular weight (~440kDa) glycoprotein of the extracellular matrix that binds to membrane-spanning receptor proteins called integrins. Fibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin. Fibronectin has numerous functions. For example, it involved in cell adhesion, cell motility, opsonization, wound healing, maintenance of cell shape, and so on. Besides, Decorin (DCN) has been identified as an interactor of FN, thus a binding ELISA assay was conducted to detect the interaction of recombinant human FN and recombinant human DCN. Briefly, FN were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to DCN-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-FN pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of of FN and DCN was shown in Figure 1, and this effect was in a dose dependent manner.

As FN has the function of cell adhesion, we measure the activity of FN by the ability of the immobilized protein to support the adhesion of NIH-3T3 mouse embryonic fibroblast cells. When 5 x 104 cells/well are added to recombinat human FN coated plates (10 µg/mL with 100 µL/well), approximately 70-80% will adhere after 3 hour incubation at 37℃. The adhesion of NIH-3T3 after 3 hour incubation at 37℃ observed by inverted microscope was shown in Figure 2.

USAGE

Reconstitute in ddH2O to a concentration of 0.1-0.5 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
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Investigative Ophthalmology & Visual Science Inhibition by Female Sex Hormones of Collagen Gel Contraction Mediated by Retinal Pigment Epithelial Cells Pubmed: 24609629
PLoS One. Mesenchymal stromal cell proliferation, gene expression and protein production in human platelet-rich plasma-supplemented media Pubmed:Pmc4130592
Connective Tissue Research Metabolic and cytoprotective effects of in vivo peri-patellar hyaluronic acid injections in cultured tenocytes Pubmed:25333747
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journal of perinatal medicine Maternal endothelial damage as a disorder shared by early preeclampsia, late preeclampsia and intrauterine growth restriction. pubmed:27865093
Chinese Journal of Contemporary Neurology and Neurosurgery Effect of panax notoginseng saponins on efficacy and hemorrhagic transformation of rt-PA intravenous thrombolysis in patients with acute ischemic stroke Article_en:CJFDTotal-XDJB201611014
Scientific Reports Serum fbronectin distinguishes the early stages of hepatocellular carcinoma pubmed:28842594
Molecular Medicine Reports Pigment epithelium‑derived factor protects human glomerular mesangial cells from diabetes via NOXO1‑iNOS suppression 10.3892/mmr.2017.7563
Cell Death & Disease Tenomodulin is essential for prevention of adipocyte accumulation and fibrovascular scar formation during early tendon healing. pubmed:29022912
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Biomedicine & Pharmacotherapy Corn silk (Zea mays L.), a source of natural antioxidants with α-amylase, α-glucosidase, advanced glycation and diabetic nephropathy inhibitory activities Pubmed: 30530231
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EBioMedicine Three kinds of corneal host cells contribute differently to corneal neovascularization
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International Journal of Women¡¯s Health and Reproduction Sciences Are the Circulating Levels of Copeptin and Fibronectin Dysregulated in Preeclamptic South African Black Women?
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IBRO Neuroscience Reports Early mobilization in spinal cord injury promotes changes in microglial dynamics and recovery of motor function Pubmed:35586775
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