Description
For Samples: Cell culture fluid & body fluid & tissue homogenate Serum or blood plasma
Intended Uses: This FIX 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 FIX in the sample, this FIX 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 FIX 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 FIX utilizes a multiclonal anti-FIX antibody and an FIX-HRP conjugate. The assay sample and buffer are incubated together with FIX-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 FIX concentration since FIX from samples and FIX-HRP conjugate compete for the anti-FIX antibody binding site. Since the number of sites is limited, as more sites are occupied by FIX from the sample, fewer sites are left to bind FIX-HRP conjugate. Standards of known FIX 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 FIX. The FIX concentration in each sample is interpolated from this standard curve