Alpha-Fetoprotein, ELISA Kit

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Catalog Number: 40-052-115007

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Product Name: Alpha-Fetoprotein

Sample Type: Serum

# of Samples: 1X96 Assays

Intended Use: For the quantitative determination of the Cancer Antigen AFP concentration in human serum.

Introduction: Alpha-fetoprotein (AFP) is a glycoprotein with a molecular weigh of approximately 70,000 daltons. AFP is normally produced during fetal and neonatal development by the liver, yolksac, and in small concentrations by the gastrointestinal tract. After birth, serum AFP concentrations decrease rapidly, and by the second year of life and thereafter only trace amounts are normally detected in serum.
Elevation of serum AFP to abnormally high values occurs in several malignant diseases, most notably nonseminomatous testicular cancer and primary hepatocellular carcinoma. In the case of nonseminomatous testicular cancer, a direct relationship has been observed between the incidence of elevated AFP levels and the stage of disease. Elevated AFP levels have also been observed in samples with seminoma with nonseminomatous elements, but not in samples with pure seminoma.
In addition, elevated serum AFP concentrations have been measured in samples with other noncancerous diseases, including ataxia telangiectasia, hereditary tyrosinemia, neonatal hyperbilirubinemia, acute viral hepatitis, chronic active hepatitis, and cirrhosis. Elevated serum AFP concentrations are also observed in pregnant women. Therefore, AFP measurements are not recommended for use as a screening procedure to detect the presence of cancer in the general population.

Principle of the test: The AFP ELISA test is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a rabbit anti-AFP antibody directed against intact AFP for solid phase immobilization (on the microtiter wells). A monoclonal anti-AFP antibody conjugated to horseradish peroxidase (HRP) is in the antibody-enzyme conjugate solution. The test sample is allowed to react first with the immobilized rabbit antibody for 30 minutes. The wells are washed to remove any unbound antigen. The monoclonal-HRP conjugate is then reacted with the immobilized antigen for 30 minutes at room temperature resulting in the AFP molecules being sandwiched between the solid phase and enzyme-linked antibodies.
The wells are washed with water to remove unbound labeled antibodies. A solution of TMB Reagent is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of Stop Solution changing the color to yellow. The concentration of AFP is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.

Storage and Stability: 1. Store the unopened kit at 2-8C upon receipt and when it is not in use, until the expiration shown on the kit label. Refer to the package label for the expiration date.
2. Keep microtiter plate in a sealed bag with desiccant to minimize exposure to damp air.

Limitation of the Test: 1. Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the package insert instructions and with adherence to good laboratory practice.
2. This product is not intended to be used for the risk calculation of trisomy 21.
3. Serum samples demonstrating gross lipemia, gross hemolysis, or turbidity should not be used with this test.
4. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.

Product Name: Alpha-Fetoprotein

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