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
American Journal of Physiology - Renal Physiology Advanced oxidation protein products induce mesangial cell perturbation through PKC-dependent activation of NADPH oxidase PubMed: 19019916
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Molecules. Identification of compounds from the water soluble extract of Cinnamomum cassia barks and their inhibitory effects against high-glucose-induced mesangial cells. Pubmed: 24013407
Journal of Ethnopharmacology Bioactive compounds from Cornus officinalis fruits and their effects on diabetic nephropathy ScienceDirect: S0378874114002414
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
Laryngoscope. Establishing principles of macromolecular crowding for in vitro fibrosis research of the vocal fold lamina propria Pubmed:25545625
Biomedical Materials In vivo bioengineered ovarian tumors based on collagen, matrigel, alginate and agarose hydrogels: a comparative study Pubmed:25634132
Biomedical Materials In vivo PubMed: 25634132
American Journal of Medical and Biological Research Impact of Nigella Sativa, Omega-3 Fatty Acids and Chromium Picolinate onNF-κB/leptin-insulin Axisin Obese Subjects with Non-alcoholic Fatty Liver Disease Ajmbr: 3
Food Chem. Comparative proteomics of milk fat globule membrane in goat colostrum and mature milk Pubmed:27173528
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
Journal of Cellular Biochemistry Effects of panax notoginseng saponin on the pathological ultrastructure and serum IL‐6 and IL‐8 in pulmonary fibrosis in rabbits Pubmed:29932250
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
Advanced Healthcare Materials Impact of Hydrogel Elasticity and Adherence on Osteosarcoma Cells and Osteoblasts Pubmed: 30838809
Inflammation TNF-α Regulates ITGβ1 and SYND4 Expression in Nucleus Pulposus Cells: Activation of FAK/PI3K Signaling Pubmed: 31111299
EBioMedicine Three kinds of corneal host cells contribute differently to corneal neovascularization
Renal Failure Effects of autophagy on macrophage adhesion and migration in diabetic nephropathy Pubmed: 31352855
Journal of Biological Chemistry Growth hormone induces Notch1 signaling in podocytes and contributes to proteinuria in diabetic nephropathy Pubmed: 31511328
Neurotherapeutics Fabrication and Evaluation of a Xenogeneic Decellularized Nerve-Derived Material: Preclinical Studies of a New Strategy for Nerve Repair Pubmed: 31758411
FASEB Journal Molecular hydrogen regulates PTEN‐AKT‐mTOR signaling via ROS to alleviate peritoneal dialysis‐related peritoneal fibrosis Pubmed: 31930571
Journal of Clinical Medicine Damage-Associated Molecular Patterns and Th-Cell-Related Cytokines Released after Progressive Effort Pubmed: 32210109
International Journal of Women¡¯s Health and Reproduction Sciences Are the Circulating Levels of Copeptin and Fibronectin Dysregulated in Preeclamptic South African Black Women?
Advanced glycation end-products associate with podocytopathy in type II diabetic patients
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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