| Field | Specification |
|---|---|
| Target | |
| Alternative names | Fibronectin;FN;Cold-insoluble globulin;CIG;Anastellin;Ugl-Y1;Ugl-Y2;Ugl-Y3;FN1;FN; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Target protein isolated from human plasma |
| Expression system | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human fibronectin (gene FN1) in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Strips are pre-coated with a capture antibody. After the sample is added, a biotinylated detection antibody binds the captured protein, and an avidin–biotin–peroxidase complex supplies the enzyme. TMB develops color in proportion to the bound target; the reaction is stopped and read at 450 nm, and concentrations are read from the standard curve.
Fibronectin (FN1) is a large extracellular matrix glycoprotein that binds integrins, collagen, fibrin and heparin. It exists as a soluble plasma form and as insoluble fibrils in tissues, and it supports cell adhesion, migration, wound healing and embryonic development. Fibronectin is a common matrix marker in studies of fibrosis and tissue repair.
Kit Components
| Anti-Human FN1 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human FN1 Standard | 2 vials, 10 ng/tube |
| Human FN1 Biotinylated antibody (100x) | 100 µL |
| Avidin-Biotin-Peroxidase Complex (100x) | 100 µL |
| Sample Diluent | 30 mL |
| Antibody Diluent | 12 mL |
| Avidin-Biotin-Peroxidase Diluent | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Wash Buffer (25x) | 20 mL |
| Adhesive plate sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; incubator; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution.
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin, EDTA, citrate) |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.6–5.7% (3 samples, n = 16 each) |
| Inter-assay CV | 5.7–7.0% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:10000. In the supplier’s tests, serum and plasma samples measured about 15 µg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.021; 156 = 0.068; 312 = 0.112; 625 = 0.197; 1250 = 0.336; 2500 = 0.605; 5000 = 1.115; 10000 = 1.83. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 156 | 312 | 625 | 1250 | 2500 | 5000 | 10000 |
| O.D. | 0.021 | 0.068 | 0.112 | 0.197 | 0.336 | 0.605 | 1.115 | 1.83 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 312 | 1118 | 5412 | 341 | 1105 | 4997 |
| Standard deviation | 14.35 | 65.76 | 308.48 | 19.43 | 76.24 | 349.79 |
| CV (%) | 4.6% | 5.7% | 5.7% | 5.7% | 6.9% | 7% |
Kit components
Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x)|100ul Avidin-Biotin-Peroxidase Complex (100x)|100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB)|10ml Stop Solution|10ml Wash Buffer (25x)|20ml Adhesive plate sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
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