|The AFP afp (Catalog #MBS590043) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase.
The AFP afp product has the following accession number(s) (GI #178236) (NCBI Accession #AAB58754.1). Researchers may be interested in using Bioinformatics databases such as those available at The National Center for Biotechnology Information (NCBI) website for more information about accession numbers and the proteins they represent. Even researchers unfamiliar with bioinformatics databases will find the NCBI databases to be quite user friendly and useful.
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Please refer to the product datasheet for known applications of a given elisa kit. We\'ve tested the Alpha Fetoprotein (AFP) with the following immunoassay(s):
Typical Testing Data/Standard Curve (for reference only)
Principle of the assay: This AFP enzyme-linked immunosorbent assay (ELISA) applies a technique called quantitative sandwich immunoassay. The microtiter plate provided in this 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. 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 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.
Background: Alpha-fetoprotein (AFP) is a glycoprotein with a molecular mass of 65,000 Daltons. It is normally produced in large amounts by liver, the visceral endoderm of foetus. Alpha-fetoprotein level increases progressively and reaches a peak at the 30th week in the maternal serum. Thereafter, it decreases gradually and reduces to trace amount with a couple of years after birth. Human AFP gene is located at the chromosomal location 4q25. The expression of AFP is regulated by a large promoter P1 and three distant enhancers. AFP expression abnormality is a relatively common genetic disorder that affects intellectual development and causes other health problems. Increased AFP expression is observed in adults that developed metastasis of liver and other organs. AFP is thought to be a foetal form of serum albumin and exists in monomeric, dimeric and trimeric forms. It binds to copper, nickel, fatty acids and bilirubin. The exact function of human AFP is still under investigation. Several hypotheses have been proposed for the physiological function: regulation of cell growth, sexual differentiation, transportation of metals and other substances, interaction with cytoplasmic chaperone proteins, and protecting foetus against the maternal immunity. To cover the very large range of concentration associated with the great variety of processes, the current immunoassay is a simple tool for the detection of AFP in the range 0 - 400ng/ml.