< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine Mouse alpha -Fetoprotein/AFP Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse AFP levels in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse AFP and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse AFP accurately. Results obtained using natural mouse AFP showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse AFP.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
The recovery of mouse AFP spiked to levels throughout the range of the assay was evaluated.
Average % Recovery
Cell Culture Media (n=4)
To assess the linearity of the assay, samples containing high concentrations of mouse AFP were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
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.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
100 µL Conjugate
Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
Aspirate and wash 4 times.
100 µL Substrate Solution
Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
100 µL Stop Solution
Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
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