Description
For Samples: Cell culture fluid & body fluid & tissue homogenate Serum or blood plasma
Intended Uses: This AOPP ELISA kit is intended for laboratory research use only and not for use in diagnostic or therapeutic procedures. The stop solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of AOPP in the sample, this AOPP ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus AOPP concentration. The concentration of in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Principle of the Assay: The coated well immunoenzymatic assay for the quantitative measurement of AOPP utilizes a multiclonal anti-AOPP antibody and an AOPP-HRP conjugate. The assay sample and buffer are incubated together with AOPP-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the AOPP concentration since AOPP from samples and AOPP-HRP conjugate compete for the anti-AOPP antibody binding site. Since the number of sites is limited, as more sites are occupied by AOPP from the sample, fewer sites are left to bind AOPP-HRP conjugate. Standards of known AOPP concentrations are run concurrently with the samples being assayed and a standard curve is plotted relating the intensity of the color (Optical Density) to the concentration of AOPP. The AOPP concentration in each sample is interpolated from this standard curve