| Field | Specification |
|---|---|
| Target | |
| Alternative names | Epidermal growth factor receptor;2.7.10.1;Proto-oncogene c-ErbB-1;Receptor tyrosine-protein kinase erbB-1;EGFR;ERBB, ERBB1, HER1; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: M1-S645 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Milk |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human epidermal growth factor receptor (gene EGFR) in cell culture supernatants, serum, plasma (heparin, EDTA) and human milk. 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.
Epidermal growth factor receptor (EGFR, also called ErbB1 or HER1) is a receptor tyrosine kinase activated by EGF, TGF-α and related ligands. Ligand binding triggers receptor dimerization and activation of RAS/MAPK, PI3K/AKT and other pathways that drive proliferation and survival, and EGFR is a major target in cancer research.
Kit Components
| Anti-Human EGFR Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human EGFR Standard | 2 vials, 10 ng/tube |
| Human EGFR 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, plasma (heparin, EDTA) and human milk |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <1 pg/mL |
| Standard | Recombinant human EGFR (Met1–Ser645), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.7–7.4% (3 samples, n = 16 each) |
| Inter-assay CV | 5.9–7.5% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:20. In the supplier’s tests, serum and plasma samples measured 41–93 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.033; 156 = 0.099; 312 = 0.149; 625 = 0.246; 1250 = 0.431; 2500 = 0.735; 5000 = 1.26; 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. | 0.033 | 0.099 | 0.149 | 0.246 | 0.431 | 0.735 | 1.26 | 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) | 289 | 1634 | 5526 | 313 | 1731 | 5572 |
| Standard deviation | 13.58 | 81.7 | 408.92 | 18.78 | 102.12 | 417.9 |
| CV (%) | 4.7% | 5% | 7.4% | 6% | 5.9% | 7.5% |
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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