| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | FETA; HPAFP; OTTHUMP00000160480|alpha-1-fetoprotein|alpha-fetoglobulin |
| Product Type | |
| Sensitivity | |
| UniProt # |
Scientific Background
AFP is a glycoprotein with a molecular weight of between 65,000 and 70,000 daltons including 4% of carbohydrate . During fetal development, AFP maintains high levels in the serum and drops to very low levels throughout the remainder of life. AFP is elevated in the malignant diseases of hepatocellular, testicular nonseminomatous origin, and occasionally of other entodermal origin. AFP may be slightly elevated or persisted in the patients with large hepatic metastases or viral hepatitis. AFP measurement is widely accepted as tumor marker and for monitoring the therapeutic effectiveness of hepato.
Assay Principle
This assay employs a two-site sandwich ELISA to quantitate AFP in samples. An antibody specific for AFP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyAFP present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for AFP is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of AFP bound in the initial step. The color development is stopped and the intensity of the color is measured.
Performance Specifications
| Sensitivity | 4 ng/mL |
|---|---|
| Detection Range | 10-400 ng/mL |
| Total Assay Time | 3.5–5 hours |
| Compatible Sample Types | serum, plasma, cell culture supernatant, tissue homogenate |
| Species Reactivity | Human |
| Detection Method | Colorimetric (TMB/HRP) |
| Storage | 4°C (short-term); -20°C (long-term) |
✓ Research-Grade Validation
Safety & Regulatory
Handle reagents in accordance with institutional biosafety guidelines. Refer to the Safety Data Sheet (SDS) for complete hazard and handling information. Contains components that may require special disposal procedures per local regulations.
This kit is validated for use with serum, plasma, cell culture supernatant, tissue homogenate. For unlisted matrices (e.g., tissue lysate, urine), perform a spike-and-recovery experiment to confirm assay performance before generating reportable data. Sample dilution in the kit's provided diluent is recommended to minimize matrix interference.
The minimum detectable concentration (sensitivity) of this kit is 4 ng/mL. Values below this threshold should be reported as below the limit of detection (<LOD) and should not be extrapolated from the standard curve.
The total assay time from sample addition to absorbance reading is approximately 3.5–5 hours, including all incubation, wash, and substrate steps. Hands-on time is typically 1–2 hours; most steps involve passive plate incubation. Plan the assay as a single uninterrupted session for best results.
Standard components of this Sandwich ELISA Kit typically include: pre-coated microplate (96-well strip format), lyophilized or liquid recombinant AFP standard, detection antibody, streptavidin-HRP conjugate, TMB substrate, stop solution, wash buffer concentrate, and sample/standard diluent. Refer to the kit insert or datasheet for the exact component list and storage requirements.
This kit uses colorimetric (TMB/HRP) detection and requires a standard microplate absorbance reader capable of measuring at 450 nm. A reference wavelength of 570 nm or 630 nm is recommended to reduce background. No specialized fluorescence or luminescence reader is needed. Ensure the instrument is calibrated and the plate is clean and free of condensation before reading.
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