| Field | Specification |
|---|---|
| Target | |
| Alternative names | Fibronectin; FN; Anastellin; Fn1 |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Target protein isolated from mouse 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 mouse 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-Mouse Fn1 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Mouse Fn1 Standard | 2 vials, 10 ng/tube |
| Mouse 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 | Mouse |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin, EDTA, citrate) |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <15 pg/mL |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 6.5–7.5% (3 samples, n = 16 each) |
| Inter-assay CV | 7.1–8.7% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:400000. In the supplier’s tests, serum and plasma samples measured about 500 µg/mL.
Example standard curve (OD450, pg/mL): 0 = 0.034; 156 = 0.092; 312 = 0.14; 625 = 0.22; 1250 = 0.397; 2500 = 0.691; 5000 = 1.251; 10000 = 1.91. 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. | 0.034 | 0.092 | 0.14 | 0.22 | 0.397 | 0.691 | 1.251 | 1.91 |
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) | 206 | 1683 | 4323 | 219 | 1843 | 3947 |
| Standard deviation | 13.39 | 126.22 | 293.96 | 15.54 | 160.34 | 335.49 |
| CV (%) | 6.5% | 7.5% | 6.8% | 7.1% | 8.7% | 8.5% |
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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