Description
The competitive Nitrotyrosine ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the inhibition principle with a working time of 3½ hours.
The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are pre-incubated with biotinylated tracer antibody in U-shape microtiter plate. Pre-incubated samples and standards with the biotinylated tracer antibodies are incubated on Nitrotyrosine-HSA (NO-HSA) coated strips. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer.
A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the known standards of the NO-HSA standards (log). The nitrotyrosine concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.