Immunoassay for detection of Polybrominated Diphenyl Ether (PBDEs) in water and soil.
Immunoassay for detection of Polybrominated Diphenyl Ether (PBDEs). Please refer to the attached specific procedures for water (groundwater, surface water, well water, effluent), and soil. Application procedures for other sample matrices can be obtained from Abraxis. The PBDE Microtiter Plate Kit applies the principles of enzyme linked immunosorbent assay (ELISA) to the determination of PBDE. In the assay system, standards or samples are added, along with an antibody specific to PBDE, to microtiter wells coated with Goat Anti-Rabbit Antibody and incubated for thirty (30) minutes. The PBDE enzyme conjugate is then added. At this point, a competitive reaction occurs between the PBDE, which may be in the sample, and the enzyme-labeled PBDE analog for the antibody binding sites on the microtiter well. The reaction is allowed to continue for thirty (30) minutes. After a washing step, the presence of PBDE is detected by adding the Color Solution, which contains the enzyme substrate (hydrogen peroxide) and the chromogen (3,3’,5,5’-tetramethylbenzidine). The enzyme-labeled PBDE bound to the PBDE antibody catalyzes the conversion of the substrate/chromogen mixture to a colored product. The color reaction is stopped and stabilized after a twenty (20) minute incubation period by the addition of diluted acid (stopping solution). The color is then evaluated using an ELISA reader. A dose response curve of absorbance vs. concentration is generated using results obtained from the standards. The concentration of PBDE present in the samples is determined directly from this curve. Since the labeled PBDE (conjugate) was in competition with the unlabeled PBDE (sample) for the antibody sites, the intensity of the color developed is inversely proportional to the concentration of Triclosan present in the sample.