| Field | Specification |
|---|---|
| Target | |
| Alternative names | Fibronectin;FN;Anastellin;Fn1; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Target protein isolated from rat 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 one-step sandwich ELISA quantifies rat fibronectin (gene Fn1) in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). The strips are pre-coated with a capture antibody; sample and an HRP-linked detection antibody are incubated together, so the assay needs fewer wash and incubation steps than a standard sandwich ELISA (the supplier markets the Quick format as a 90-minute assay). TMB develops color in proportion to the bound target, and the plate is read at 450 nm.
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
| 96-well plate precoated with anti-Rat Fn1 antibody | 1 |
|---|---|
| lyophilized recombinant Rat Fn1 standard | 10 ng/tube |
| Biotinylated anti-Rat Fn1 detection antibody (50x) | 100 µL |
| Sample diluent | 30 mL |
| Antibody diluent | 12 mL |
| Wash Buffer (25x) | 20 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Plate Sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; 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 | Rat |
|---|---|
| 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 | 4.4–6.8% (3 samples, n = 16 each) |
| Inter-assay CV | 5.4–8.3% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.012; 156 = 0.111; 313 = 0.208; 625 = 0.381; 125000 = 0.668; 25000 = 1.201; 5000 = 1.952; 10000 = 2.461. 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 | 313 | 625 | 125000 | 25000 | 5000 | 10000 |
| O.D. | 0.012 | 0.111 | 0.208 | 0.381 | 0.668 | 1.201 | 1.952 | 2.461 |
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) | 342 | 1852 | 4326 | 361 | 1834 | 4010 |
| Standard deviation | 23.25 | 85.4%9 | 190.34 | 29.96 | 102.7 | 216.54 |
| CV (%) | 6.8% | 4.6% | 4.4% | 8.3% | 5.6% | 5.4% |
Kit components
Description|Quantity 96-well plate|1 lyophilized Standard| Biotinylated detection antibody (50x)|100 ul Avidin-Biotin-Peroxidase Complex (30x)|400 µl Sample diluent|30 ml Antibody diluent|12 ml Avidin-Biotin-Peroxidase Diluent|12 ml Wash Buffer (25x)|20 ml Color Developing Reagent (TMB)|10 ml Stop Solution|10 ml Plate Sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- 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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