Alkyl Ethoxylate (AE), ELISA, 96-test
Immunoassay for the quantitative and sensitive detection of Alkyl Ethoxylate (AE). Kit Principle Competitive Reaction The test is based on the recognition of AE by specific monoclonal antibodies. AE present in the sample and an AE-enzyme conjugate (i.e. AE labeled with a coloring enzyme) are premixed and added into each well of a microplate, and allowed to compete for limited number of binding sites of specific antibodies immobilized on the surface of the wells. When the AE concentration is higher relative to the enzyme conjugate, the AE will predominantly bind the antibody and vice versa. Chromogenic Reaction Unbound AE and excess AE-enzyme conjugates are washed out. The presence of AE is detected by adding a chromogenic substrate. The enzyme-labeled AE bound to the AE antibody in the plate, catalyzes the conversion of the substrate to a colored product. After an incubation period, the reaction is stopped by the addition of a diluted acid. The higher the AE concentration in a sample, for example, leads to less antigen-enzyme conjugate bound to the antibody binding sites in a microplate well, generating a lighter color, i.e. lower absorbance. Quantitative Analysis The standard curve, a dose-response curve obtained from known concentrations of AE standards, is determined from the absorbance at450nm . The AE concentration in each sam ple is accurately calculated by interpolating using the absorbance values from the standard curve. Kit Features -The monoclonal antibody binds exclusively with C10-12 AEs regardless of ethylene oxide (EO) chain length (n=1-25) and does not show cross reaction with other chemicals of similar structures. A monoclonal antibody is uniform in quality, generating very little lot-to-lot variation. -The quantitative analysis ranges from 0.02mg/L and 1.0mg/L. A simple concentration protocol based on solid phase extraction is available to determine much lower concentration. -The ELISA measurement is highly reproducible; the coefficient of variation (CV) is mostly under 10%. -The assay requires less amount of harmful solvent than instrument analyses. -With ease of handling, the total time for measurement is only 2.5 hours. -The kit, a 96-well microplate format, enables simultaneous measurement of multiple samples at more reasonable cost.