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|Method & Line||Sample & Target||Product||Package Info|
|MicroELISA||Serum||EIAgen GI-Marker CA 19-9 Kit||Tests per Package: 96|
|EIAgen||The EIAgen GI-Marker CA 19-9 Kit has been designed for the quantitative determination of Cancer Antigen CA 19-9 (CA 19-9) Concentration in Human Serum.||Code: LI4040F1||Package: 1 Microplate|
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The EIAgen GI-Marker CA 19-9 Kit has been designed for the quantitative determination of Cancer Antigen CA 19-9 (CA 19-9) Concentration in Human Serum.
A mucin type Sialyl Lewis Antigens group of glycoproteins (SLA) such as CA 19-9, 19-5 have been recognized as circulating cancer associated antigens for gastrointestinal cancer. The discovery of a monoclonal antibody clone (1116NS 19-9), which exhibited selective reactivity with human gastrointestinal carcinomas through the recognition of a carbohydrate determinant (CA 19-9) defined as a sialyl lacto-N-flucopenrose II, resulted in the successful purification and thus, determination of human gastrointestinal tumor associated glycoprotein antigen expressing CA 19-9 from colorectal carcinoma cell lines. Recently reports indicate that serum CA 19-9 level is frequently elevated in the circulation of patients with various gastrointestinal malignancies, such as pancreatic, colorectal, gastric and hepatic carcinomas. Together with CEA elevated CA 19-9 is suggestive of gallbladder disease. The tumor associated antigen may also be associated in some malignant conditions. Research studies demonstrate that serum CA 19-9 values may have utility in monitoring subjects with the above mentioned diagnosed malignancies.
In this method, CA19-9 calibrator, patient specimen or control is first added to a streptavidin coated well. Biotinylated monoclonal antibody (specific for CA19-9) is added and the reactants mixed. Reaction between the CA19-9 antibodies and native CA19-9 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 monoclonal antibody specific to CA19-9 is added to the wells. The enzyme labeled antibody binds to the CA19-9 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 CA19-9 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-CA19-9 antibody.
Upon mixing monoclonal biotinylated antibody, and a serum containing the native antigen, 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 LI4040F1).
Bring reagents to room temperature before use.
CA 19-9 Biotin
Wash Buffer Conc. 50X
Number of tests