| Field | Specification |
|---|---|
| Target | |
| Alternative names | Vascular endothelial growth factor A, long form; L-VEGF; Vascular permeability factor; VPF; VEGFA; VEGF |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: A27-R191 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This single-incubation sandwich ELISA quantifies human vascular endothelial growth factor A, long form (gene VEGFA) in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). The strips are coated with an anti-tag antibody. Samples and standards are added to the wells together with the kit’s antibody cocktail (Antibody A and Antibody B); the resulting antibody–target complex is held on the plate through the tag. After washing, TMB substrate is added and the HRP label on the detection antibody converts it to a colored product in proportion to the bound target. The reaction is stopped and the plate is read at 450 nm. The supplier markets the Quick format as a 90-minute assay.
VEGF-A is the main vascular endothelial growth factor that drives angiogenesis. Produced in response to hypoxia, it binds VEGFR1 and VEGFR2 on endothelial cells to promote their proliferation, migration and survival and to increase vascular permeability. VEGF-A is widely studied in vascular development, wound healing, eye disease and tumor angiogenesis.
Kit Components
| Anti-tag Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Human VEGFA Standard | 2 × 10 ng/tube |
| Human VEGFA Antibody A | 3 mL |
| Human VEGFA Antibody B | 3 mL |
| Sample Diluent | 15 mL |
| TBS-T Wash Buffer (25x) | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Adhesive Plate Sealers | 2 |
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; horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3 mm.
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin, EDTA, citrate) |
| Assay range | 31.2 pg/mL–2,000 pg/mL (human serum,plasma), 15.6 pg/mL–1,000 pg/mL (cell culture supernates) |
| Sensitivity | <1 pg/mL |
| Standard | Recombinant human VEGFA (Ala27–Arg191), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 7.8–9.5% (3 samples, n = 16 each) |
| Inter-assay CV | 7.4–9.4% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.130; 31.2 = 0.338; 62.5 = 0.523; 125 = 0.912; 250 = 1.480; 500 = 2.228; 1000 = 2.473; 2000 = 2.536. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months. Ships on gel ice and tolerates up to 3 days at room temperature; refrigerate on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.130 | 0.338 | 0.523 | 0.912 | 1.480 | 2.228 | 2.473 | 2.536 |
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) | 77 | 280 | 953 | 72 | 263 | 988 |
| Standard deviation | 7.32 | 21.84 | 89.58 | 5.54 | 24.72 | 73.11 |
| CV (%) | 9.5% | 7.8% | 9.4% | 7.7% | 9.4% | 7.4% |
Kit components
Description|Quantity|Volume Anti-tag Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 Antibody A|1|3ml Antibody B|1|3ml Sample Diluent|1|15ml TBS-T Wash Buffer (25x)|1|12ml Color Developing Reagent (TMB)|1|10ml Stop Solution|1|10ml Adhesive Plate Sealers|2|PieceMaterials 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.
- Horizontal orbital microplate shaker capable of maintaining a speed of 500 rpm, amplitude 3mm.
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