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This kit is comprised of microplate coated with AIV (H5N1) antigen, enzyme-conjugated second antibody and other reagents. It utilizes the principles of ELISA to detect antibody specific to AIV (H5N1) in chicken serum and plasma. Control serum and diluted serum samples are added into the microplate during experiment. After incubation, if the sample contains antibody specific to AIV (H5N1), the antibody will bind with the antigen coated in the microplate. The microplate should be washed with wash solution to remove unbound antibody and other components. The enzyme- conjugated second antibody is added into the washed microplate and it will specifically bind with the antibody/antigen complex on the microplate. The microplate should be washed once again to remove unbound enzyme conjugate. Then TMB substrate solution is added into wells to react with enzyme and appears blue in color. The enzyme-substrate reaction is terminated by the addition of the stop solution, the color changes from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm and proportional to the concentration of specific antibody.
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