| Field | Specification |
|---|---|
| Target | |
| Alternative names | Alpha-fetoprotein;Alpha-1-fetoprotein;Alpha-fetoglobulin;AFP;HPAFP; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: R19-V609 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Cell Lysate, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This one-step sandwich ELISA quantifies human alpha-fetoprotein (gene AFP) in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). 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.
Alpha-fetoprotein (AFP) is the major serum protein of the fetus, produced mainly by the fetal liver and yolk sac and related in structure to albumin. It binds fatty acids, metal ions and bilirubin, and its levels are widely measured in studies of fetal development and liver tumors such as hepatocellular carcinoma.
Kit Components
| Anti-Human AFP Pre-coated 96-well Strip Microplate | 12 strips of 8 wells |
|---|---|
| Human AFP Standard | 2 × 5 ng/tube |
| HRP Linked Anti-Human AFP Antibody | 6 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): reader capable of reading absorbance at 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.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 78 pg/mL–5,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human AFP (Arg19–Val609), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
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 | 78 | 156 | 312 | 625 | 1250 | 2500 | 5000 |
| O.D. | 0.003 | 0.137 | 0.248 | 0.486 | 0.777 | 1.260 | 1.704 | 1.925 |
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) | 98 | 643 | 2190 | 98 | 654 | 1986 |
| Standard deviation | 4.56 | 47.58 | 100.74 | 5.58 | 56.24 | 129.09 |
| CV (%) | 4.8% | 7.4% | 4.6% | 5.7% | 8.6% | 6.5% |
Kit components
Description|Quantity|Volume Pre-coated 96-well Strip Microplate|1|12 strips of 8 wells Standard||2 HRP-Linked Antibody|1|6ml 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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