Method Description

The assay is performed using a Beckman Coulter Unicel DXI 800. The Access Inhibin A assay is a sequential 2-step immunoenzymatic ("sandwich'') assay. Sample is added to a reaction vessel and incubated with paramagnetic particles coupled with anti-inhibin A monoclonal antibody. Excess sample and reagents are removed, and anti-inhibin A monoclonal antibody-alkaline phosphatase conjugate is then added to a reaction mixture. After incubation, unbound materials are washed away. Antibody-analyte complex is detected by addition of the chemiluminescent substrate. The light production is directly proportional to the concentration of inhibin A in the sample.(Package insert: Access Inhibin A. Beckman Coulter Inc; 10/2023)

The ultra-sensitive inhibin B enzyme-linked immunosorbent assay is a quantitative three-step sandwich type immunoassay. Sample is incubated in wells that have been coated with inhibin B antibody. After incubation and washing, the wells are incubated with biotinylated inhibin B antibody. After a second incubation and washing step, the wells are incubated with streptavidin horseradish peroxidase conjugate. After the third incubation and washing step, the wells are incubated with substrate solution. After incubation, an acidic stopping solution is added. Antibody-analyte complex is detected by dual wavelength absorbance measurement at 450 nm as the primary test filter and 620 nm as the reference filter. The absorbance measured is directly proportional to the concentration of inhibin B in the samples and calibrators.(Unpublished Mayo method)