The ADMA Mouse/Rat ELISA Kit is intended for the quantitative determination of asymmetric dimethyl-L-arginine (ADMA) in rodent EDTA plasma or serum and cell culture media.
For research use only. Not for use in diagnostic procedures.
For Laboratory Professional Use Only.
Developed by Immundiagnostik AG.
The ADMA Mouse/Rat ELISA is based on the method of competitive enzyme-linked immunoassays. The sample preparation includes the addition of a derivatization reagent for ADMA derivatization. Afterward, the treated samples and the polyclonal ADMA-antiserum are incubated in the wells of a microtiter plate coated with ADMA-derivative (tracer).
During the first incubation period, the target ADMA in the sample competes with the tracer immobilized on the wall of the microtiter wells to bind the polyclonal antibodies.
During the second incubation step, a peroxidase-conjugated antibody is added to detect the anti-ADMA antibodies. After washing away the unbound components, tetramethylbenzidine (TMB) is added as a peroxidase substrate. Finally, the enzymatic reaction is terminated by an acidic stop solution. The color changes from blue to yellow, and the absorbance is measured in the photometer at 450 nm. The intensity of the yellow color is inversely proportional to the ADMA concentration in the sample. This means a high ADMA concentration in the sample reduces the concentration of tracer-bound antibodies and lowers the photometric signal.
A dose-response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standard. The presence of ADMA in the samples is determined directly from this curve.
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