alpha-Fetoprotein (AFP), is albuminoid superfamily protein that is synthesized in the fetus primarily by the liver, yolk sac, and tissues of gastrointestinal origin. It is one of the earliest markers of the hepatocyte lineage. AFP acts as a carrier protein for steroids, bilirubin, fatty acids, retinoids, and flavonoids. In addition, it can exert immunosuppressive activity, regulate cell proliferation and apoptosis, initiate intracellular signaling, and contribute to cell invasion. Altered levels of both fetal and maternal AFP have been associated with hypothyroidism, autoimmune disorders, and heart defects. Low maternal serum AFP levels are associated with a higher incidence of Down syndrome, whereas higher levels are associated with spina bifida and anencephaly. Elevated AFP levels are also coincident with liver, stomach, and germ cell cancers.
Human alpha-Fetoprotein Quantikine ELISA Kit
R&D Systems | Catalog # DAFP00
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)
Sensitivity
0.046 ng/mL
Assay Range
0.312-20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human alpha-Fetoprotein Quantikine ELISA Kit
The Quantikine Human AFP immunoassay is a 4.5 hour solid phase ELISA designed to measure human AFP in cell culture supernates, serum, and plasma.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural human AFP
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity not tested.
Interference
Interference observed with other substances.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (ng/mL) | 2.89 | 6.02 | 12.3 | 2.65 | 5.62 | 11.7 |
| Standard Deviation | 0.109 | 0.219 | 0.441 | 0.158 | 0.261 | 0.549 |
| CV% | 3.8 | 3.6 | 3.6 | 6.0 | 4.6 | 4.7 |
EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (ng/mL) | 3.15 | 6.62 | 13.2 | 3.22 | 6.79 | 14.1 |
| Standard Deviation | 0.080 | 0.149 | 0.447 | 0.215 | 0.382 | 0.930 |
| CV% | 2.5 | 2.3 | 3.4 | 6.7 | 5.6 | 6.6 |
Recovery for Human alpha-Fetoprotein Quantikine ELISA Kit
The recovery of AFP spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 99 | 95-103 |
| EDTA Plasma (n=4) | 104 | 94-114 |
| Heparin Plasma (n=4) | 103 | 95-115 |
| Serum (n=4) | 103 | 86-115 |
Linearity
To assess the linearity of the assay, samples spiked with high concentrations of AFP were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human alpha-Fetoprotein Quantikine ELISA Kit
Human alpha-Fetoprotein/AFP ELISA Calibrator Diluent RD5-24 Standard Curve
Human alpha-Fetoprotein/AFP ELISA Calibrator Diluent RD5-33 Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: alpha-Fetoprotein/AFP
Additional alpha-Fetoprotein/AFP Products
Product Documents for Human alpha-Fetoprotein Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human alpha-Fetoprotein Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human alpha-Fetoprotein Quantikine ELISA Kit
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Protocols
View specific protocols for Human alpha-Fetoprotein Quantikine ELISA Kit (DAFP00):
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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