The Nitrotyrosine ELISA is an enzyme immunoassay intended for the quantitative determination of protein-bound nitrotyrosine in EDTA plasma, serum, and stool.

For research use only. Not for use in diagnostic procedures.

For Laboratory Professional Use Only.

This is the RUO version of Immundiagnostik part number K 7824.

Test Principle of the Nitrotyrosine ELISA

The Nitrotyrosine ELISA is designed for the quantitative determination of nitrotyrosine. It utilizes the sandwich technique with two polyclonal antibodies against nitrated proteins. Standards, controls, and diluted samples assayed for nitrotyrosine are added into the wells of a microtiter plate coated with polyclonal anti-nitrotyrosine antibody.

During the first incubation step, nitrated proteins are bound by the immobilized primary antibody. Then a peroxidase-conjugated polyclonal anti-nitrotyrosine antibody is added into each microtiter well, and a sandwich of primary antibody – nitrated protein – peroxidase-conjugate is formed. Tetramethylbenzidine is used as peroxidase substrate.

Finally, an acidic stop solution is added to terminate the reaction. The color changes from blue to yellow. The intensity of the yellow color is directly proportional to the nitrotyrosine concentration. A dose-response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standards. The presence of nitrotyrosine in the samples is determined directly from this curve.