The Experiment Principle For Elisa Kits
At first the microtiter plate provided in ELISA Kit has been precoated with an antibody specific to the target protein.
Then Standards or samples are added to the appropriate microtiter plate wells with a biotin-conjugated specific polyclonal antibody preparation, and Avidincon jugated to Horseradish Peroxidase(HRP) is added to each microplate well and incubated.
Then a TMB substrate solution is added to each well. Only those wells that contain target protein, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.
The concentration of the samples is then determined by comparing the O.D. of the samples to the standard curve.
Then Standards or samples are added to the appropriate microtiter plate wells with a biotin-conjugated specific polyclonal antibody preparation, and Avidincon jugated to Horseradish Peroxidase(HRP) is added to each microplate well and incubated.
Then a TMB substrate solution is added to each well. Only those wells that contain target protein, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm.
The concentration of the samples is then determined by comparing the O.D. of the samples to the standard curve.
