| Field | Specification |
|---|---|
| Target | |
| Alternative names | Vascular endothelial growth factor receptor 3;VEGFR-3;2.7.10.1;Fms-like tyrosine kinase 4;FLT-4;Tyrosine-protein kinase receptor FLT4;Flt4;Flt-4, Vegfr3; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: Y25D770 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies mouse vascular endothelial growth factor receptor 3 (gene Flt4) in cell culture supernatants and serum. 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.
Vascular endothelial growth factor receptor 3 (VEGFR-3, FLT4) is a receptor tyrosine kinase activated by VEGF-C and VEGF-D. It is essential for the growth of lymphatic vessels and also contributes to early blood vessel development. VEGFR-3 is a common lymphatic endothelial marker, and FLT4 mutations cause hereditary lymphedema (Milroy disease).
Kit Components
| Anti-Mouse Flt4 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Mouse Flt4 Standard | 2 vials, 10 ng/tube |
| Mouse Flt4 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 and serum |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant mouse Flt4 (Y25D770), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.9–7.8% (3 samples, n = 16 each) |
| Inter-assay CV | 7.2–9.5% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:1. In the supplier’s tests, serum samples measured about 1 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.058; 156 = 0.118; 312 = 0.179; 625 = 0.3; 1250 = 0.557; 2500 = 0.962; 5000 = 1.638; 10000 = 2.243. 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. | 0.058 | 0.118 | 0.179 | 0.3 | 0.557 | 0.962 | 1.638 | 2.243 |
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) | 287 | 1392 | 5844 | 306 | 1363 | 5406 |
| Standard deviation | 22.38 | 87.69 | 344.79 | 29.07 | 99.49 | 389.23 |
| CV (%) | 7.8% | 7.3%% | 5.9% | 9.5% | 7.3% | 7.2% |
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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Siyuan Su et al. (2022) Structure elucidation, immunomodulatory activity, antitumor activity and its molecular mechanism of a novel polysaccharide from Boletus reticulatus Schaeff. Food Science and Human Wellness. 10.1016/j.fshw.2022.07.067
Ryo Shimada et al. (2022) Gene expression in meningeal lymphatic endothelial cells following traumatic brain injury in mice. PLoS One. 10.1371/journal.pone.0273892
