Fibronectin (FN) is a large modular glycoprotein that is found as a polymeric fibrillar network in the extracellular matrix (ECM) and as soluble disulfide-linked dimeric protomers in plasma and other body fluids. Fibronectin is a ligand for many molecules including fibrin, heparin, chondroitin sulfate, collagen/gelatin, and integrins. It is involved in multiple cellular processes such as cell adhesion/migration, blood clotting, morphogenesis, tissue repair, and cell signaling. Fibronectin functions are mediated by the insoluble polymeric fibrils. Conversion of soluble fibronectin to fibronectin fibrils in the ECM is initiated by binding to cell surface integrins, resulting in exposure of cryptic epitopes necessary for polymerization.
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Human Fibronectin Quantikine ELISA Kit
R&D Systems | Catalog # DFBN10
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Cell Lysates (50 µL), Tissue Lysates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Citrate Plasma (10 µL), Saliva (17 µL), Urine (17 µL), Human Milk (10 µL)
Sensitivity
0.579 ng/mL
Assay Range
3.13-200 ng/mL (Cell Culture Supernates, Cell Lysates, Tissue Lysates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Saliva, Urine, Human Milk)
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Product Summary for Human Fibronectin Quantikine ELISA Kit
The Quantikine Human Fibronectin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Fibronectin in serum-free cell culture supernates, cell lysates, tissue lysates, serum, plasma, saliva, urine, and human milk.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
natural and recombinant human Fibronectin and Fibronectin fragments aa 1-1908, aa 607-1265, aa 627-702. Fibronectin fragment aa 631-705 cross-reacts approximately 0.415% Fibronectin fragments aa 1266-1908, aa 1722-1811, and aa 1913-2477 are not detected in this kit.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Cell Lysates, Citrate Plasma, EDTA Plasma, Heparin Plasma, Human Milk, Saliva, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (ng/mL) | 24.0 | 58.1 | 123 | 23.3 | 56.5 | 115 |
| Standard Deviation | 1.40 | 2.78 | 6.35 | 1.31 | 2.98 | 6.38 |
| CV% | 5.8 | 4.8 | 5.2 | 5.6 | 5.3 | 5.5 |
Recovery for Human Fibronectin Quantikine ELISA Kit
The recovery of human Fibronectin spiked to levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 97 | 89-115 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Fibronectin were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Scientific Data Images for Human Fibronectin Quantikine ELISA Kit
Human Fibronectin ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Fibronectin
Alternate Names
CIG, ED-B, FINC, FN1, FNZ, GFND2, LETS, SMDCF
Gene Symbol
FN1
Additional Fibronectin Products
Product Documents for Human Fibronectin Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Fibronectin Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Citations for Human Fibronectin Quantikine ELISA Kit
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Protocols
View specific protocols for Human Fibronectin Quantikine ELISA Kit (DFBN10):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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