Description
The Eagle Biosciences Human Adrenocorticotrophic Hormone (ACTH) ELISA Assay Kit is designed, developed and produced for the quantitative measurement of human ACTH in EDTA-plasma sample. The assay utilizes the two-site “sandwich” technique with selected antibodies that bind to N-terminal and C-terminal epitopes of ACTH. Assay standards, controls and patient samples are added directly to wells of a microtiter plate that is coated with antibody to the N-terminal of human ACTH. Immediately, a horseradish peroxidase (HRP) conjugated anti C-terminal of human ACTH antibody is added to each well. After the first incubation period, a “sandwich” of solid-phase monoclonal antibody - human ACTH – HRP conjugated antibody” is formed. The unbound antibodies and buffer matrix are removed in the subsequent washing step. For the detection of this immunocomplex, the well is then incubated with a substrate solution in a timed reaction and the absorbances are then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to the wall of each microtiter well is directly proportional to the amount of human ACTH in the test sample. A standard curve is generated by plotting the absorbance versus the respective human ACTH concentration for each standard on a point-to-point or 4-parameter curve fitting. The concentration of human ACTH in test samples is determined directly from this standard curve