Description
Assay standards, controls and test samples are directly added to wells of a microplate that is coated with streptavidin. Subsequently, a mixture of biotinylated GLP-1 specific antibody and a horseradish peroxidase (HRP)-conjugated GLP-1 specific antibody is added to each well. After the first incubation period, a “sandwich” immunocomplex of “Streptavidin – Biotin-Antibody – GLP-1(7-36)/(9-36) – HRP-conjugated antibody” is formed and attached to the wall of the plate. The unbound HRP-conjugated antibody is removed in a subsequent washing step. For the detection of this immunocomplex, each well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to GLP-1 (7-36)/(9-36) on the wall of the microtiter well is directly proportional to the amount of Total GLP-1 in the sample