AFP ELISA KIT
INTENDED USE
This AFP ELISA Kit is to be used for the in vitro quantitative determination of alphafetoprotein in human serum. This AFP ELISA Kit is intended FOR LABORTORY RESEARCH USE ONLY and is not for use in diagnostic or therapeutic procedures.
INTRODUCTION
Alphafetoprotein (AFP) is a glycoprotein with a molecular mass of 65000 daltons. It is normally produced in large amounts by the foetal liver. Alphafetoprotein level increases progressively and reaches a peak at the 30th week in the maternal serum, thereafter, it decreases gradually. An elevated maternal serum alphafetoprotein is associated with the neural tube defect (spina bifida) and placental abnormalities, whereas decreased AFP levels in both maternal serum and amniotic fluid are related to foetal chromosomal abnormalities (Down-syndrome). Patients affected by liver carcinoma, testicular or ovarian teratocarcinoma present very high AFP levels. Although less frequently, an increased alphafetoprotein concentration may also be observed in gastric, breast and bronchial tumors. To cover the very large range of concentration associated with the great variety of pathologic processes, the current microtiter plate immunoassay (ELISA) is a simple and powerful diagnostic tool for the detection of AFP in the range 0 – 400ng/ml.
PRINCIPLE OF THE ASSAY
AFP ELISA Kit
AFP ELISA Kit is to be used for the in vitro quantitative determination of alphafetoprotein in human serum. This AFP ELISA Kit is intended FOR LABORTORY RESEARCH USE ONLY and is not for use in diagnostic or therapeutic procedures. This AFP ELISA Kit applies a technique called quantitative sandwich immunoassay.
The microtiter plate provided in this AFP ELISA Kit has been pre-coated with a monoclonal antibody specific for AFP. Standards or samples are then added to the appropriate microtiter plate wells and incubated. AFP if present, will bind and become immobilized by the antibody pre-coated on the wells. The microtiter plate wells are thoroughly washed to remove any unbound AFP and other components of sample. In order to quantitate the amount of AFP present in the sample, a standardized preparation of horseradish peroxidase (HRP) conjugated monoclonal antibody specific for AFP is added to each well to "sandwich" the AFP immobilized during the first incubation. The microtiter plate then undergoes a second incubation. The wells are thoroughly washed to remove all unbound HRP-conjugated antibodies and a TMB (3,3'5,5' tetramethyl-benzidine) substrate solution is added to each well. The enzyme (HRP) and substrate solution are allowed to react over a short incubation period. Only those wells that contain AFP and enzyme-substrate reaction will exhibit a change in colour. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the colour change measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.
In order to measure the concentration of AFP in the samples, this AFP ELISA Kit includes a set standard .This allows the operator to produce a standard curve of Optical Density (O.D) versus AFP (ng/mL). The concentration of AFP in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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