Alpha-Fetoprotein (AFP) ELISA
The Calbiotech AFP ELISA Kit is intended for the quantitative measurement of AFP in human serum.
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 patients diagnosed with seminoma with nonseminomatous elements, but not in patients with pure seminoma.
In addition, elevated serum AFP concentrations have been measured in patients 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.
This AFP ELISA kit is a solid phase sandwich assay method, based on a streptavidin-biotin principle. The standards, samples and the biotinylated Anti-AFP antibody reagent are added into designated wells, coated with Streptavidin. Endogenous AFP in the patient’s serum binds to the antigenic site of the biotinylated Anti-AFP antibody. Simultaneously, the biotinylated antibody is immobilized onto the wells through the high affinity Streptavidin-Biotin interaction. Unbound protein and excess biotin conjugated antibody are washed off by wash buffer. Upon the addition of the Peroxidase (HRP) conjugated Anti-AFP antibody reagent, a sandwich complex is formed, the analyte of interest being in between the two highly specific antibodies, labeled with Biotin and HRP. Unbound protein excess enzyme conjugated antibody reagent is washed off by wash buffer. Upon the addition of the substrate, the intensity of color developed is directly proportional to the concentration of AFP in the samples. A standard curve is prepared relating color intensity to the concentration of the AFP.
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