CEA ELISA kit can be used for the quantitative determination of the Cancer Antigen CEA concentration in human serum.The CEA MTPL-EIA kit is based on the principle of a Solid-Phase-Enzyme-Linked immunosorbent assay. The CEA ELISA kit utilizes a monoclonal antibody directed against a distinct antigenic determinant on the intact CEA molecule is used for solid phase immobilization (on the microtiter wells). A goat anti-CEA antibody conjugated to horseradish peroxidase (HRPO) is in the antibody-enzyme conjugate solution. The test sample is allowed to react simultaneously with the two antibodies, resulting in the CEA molecules being sandwiched between the solid phase and enzyme-linked antibodies. After a 1 hour incubation at room temperature, the wells are washed with water to remove unbound labelled antibodies. A solution of H2O2/TMB is added and incubated for 20 minutes, resulting in the development of a blue colour. The colourdevelopment is stopped with the addition of 2N HCl changing the colour to yellow. The concentration of CEA is directly proportional to the colour intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.
Unopened CEA ELISA kits should be stored at 2-8°C upon receipt and the microtiter plate of the CEA ELISA kit should be kept in a sealed bag with desiccants to minimize exposure to damp air. Opened CEA ELISA kits will remain stable until the expiration date shown, provided it is stored as described above. A microtiter plate reader with a bandwidth of 10 nm or less and an optical density range of 0-2 OD or greater at 450 nm wavelength is acceptable for use in absorbance measurement.
The most complete study of CEA is a compilation of collaborative studies in which CEA value in 35,000 samples from more than 10,000 patients and controls were analyzed. Of 1425 normal persons who did not smoke, 98.7% had values less than 5.0 ng/ml. It is recommended that each laboratory establish its own normal range. The minimum detectable concentration of CEA by this assay is estimated to be 1.0 ng/ml.
The test results using the CEA ELISA kit are only valid if the test has been performed following the instructions. Moreover the user must strictly adhere to the rules of GLP (Good Laboratory Practice) or other applicable standards/laws. All standards must be found within the acceptable ranges as stated on the QC Certificate of the CEA ELISA kit. If the criteria are not met, the run is not valid and should be repeated. Each laboratory should use known samples as further controls. In case of any deviation the following technical issues should be proven: Expiration dates of (prepared) reagents, storage conditions, pipettes, devices, incubation conditions and washing methods. It is recommended to participate at appropriate quality assessment trials.
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