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|Method & Line||Sample & Target||Product||Package Info|
|MicroELISA||Serum||EIAgen BR-Marker CA 15-3 Kit||Tests per Package: 96|
|EIAgen||The EIAgen BR-Marker CA 15-3 Kit has been designed for the quantitative determination of Cancer Antigen CA 15-3 (CA 15-3) Concentration in Human Serum.||Code: LI4036F1||Package: 1 Microplate|
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The EIAgen BR-Marker CA 15-3 Kit has been designed for the quantitative determination of Cancer Antigen CA 15-3 (CA 15-3) Concentration in Human Serum.
Although multiple serum based tumor markers have been described for breast cancer, such as CA 15-3, BR 27-29, carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA), tissue polypeptide specific antigen, and HER-2 (the extracellular domain), the most widely used are CA 15-3 and CEA. CA 15-3 is considered to be one of the first circulating prognostic factors for breast cancer (1). Preoperative concentrations thus might be combined with prognostic factors for predicting outcome in patients with newly diagnosed breast cancer(2). At present the most important clinical application of CA 15-3 is in monitoring therapy in patients with advanced breast cancer that is not accessible by existing clinical or radiologic procedures(3).
The CA 15-3 assay measures the protein product of MUC1 gene. MUC1 protein is a large transmembrane glycosylated molecule containing three main domains, a large extracellular region, a membrane spanning sequence, and a cytoplasmic domain (4). Although the physiologic function of MUC1 is unclear, the glycoprotein has been implicated in cell adhesion, immunity and metastasis. Compared with healthy breast tissue, MUC1 is present in higher concentrations but less glycosylated in breast carcinoma (5-8).
In this method, a prediluted CA15-3 calibrator diluted patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal antibody (specific for CA15-3) is added and the reactants mixed. Reaction between the CA15-3 antibodies and native CA15-3 forms complex that binds with the streptavidin coated to the well. The excess serum proteins are washed away via a wash step. Another enzyme labeled antibody specific for a different epitopic recognition of CA15-3 is added to the wells. The enzyme labeled antibody binds to the CA15-3 already immobilized on the well through its binding with the biotinylated monoclonal antibody. Excess enzyme is washed off via a wash step. A color is generated by the addition of a substrate. The intensity of the color generation is directly proportional to the concentration of the CA15-3 in the sample.
The essential reagents required for an immunoenzymometric assay include high affinity and specificity antibodies (enzyme and immobilized), with different and distinct epitope recognition, in excess, and native antigen. In this procedure, the immobilization takes place during the assay at the surface of a microplate well through the interaction of streptavidin coated on the well and exogenously added biotinylated monoclonal anti-CA15-3 antibody.
Upon mixing monoclonal biotinylated antibody, and a serum containing the native antigen, a reaction results between the native antigen and the antibody, forming an antibody-antigen complex.
Simultaneously, the complex is deposited to the well through the high affinity reaction of streptavidin and biotinylated antibody.
After a suitable incubation period, the antibody-antigen bound fraction is separated from unbound antigen by decantation or aspiration. Another antibody (directed at a different epitope) labeled with an enzyme is added. Another interaction occurs to form an enzyme labeled antibody-antigen-biotinylated-antibody complex on the surface of the wells. Excess enzyme is washed off via a wash step. A suitable substrate is added to produce color measurable with the use of a microplate spectrophotometer. The enzyme activity on the well is directly proportional to the native free antigen concentration. By utilizing several different serum references of known antigen concentration, a dose response curve can be generated from which the antigen concentration of an unknown can be ascertained.
The kit contains reagents for 96 tests (code LI4036F1).
Bring reagents to room temperature before use.
CA 15-3 Biotin
Sample / Controls Diluent
Wash Buffer Conc. 50X
Number of tests