Description
The GFAP ELISA Assay Kit test is based on the principle of a solid phase enzyme-linked sandwich immunosorbent assay. The assay system utilizes a specific monoclonal antibody directed against a distinct antigenic determinant on the GFAP molecule and is coated on the microtiter wells for the solid phase immobilization of GFAP. A biotin labeled rabbit anti-GFAP antibody is used as a reporter molecule in a sandwich immunoassay and streptavidin conjugated to Horse radish peroxidase (HRP) is used as the detector molecule. The test sample (serum) is allowed to react with the capture antibody which immobilizes GFAP present in the sample. Following washing, biotinylated polyclonal reporter antibody is added to the wells resulting in the GFAP molecule being sandwiched between the solid phase and biotin-labeled antibodies. After an additional incubation with streptavidin-HRP the wells are washed, a TMB substrate solution is added and the relative absorbance units (AU) are measured spectrophotometrically using a microtiter plate reader at 450nm. The concentration of GFAP is directly proportional to the AUs of the test sample and is determined from the standard curve