Adiponectin (Rat) ELISA

This assay is a Sandwich ELISA based, sequentially, on: 1) capture of Adiponectin molecules from samples to the wells of a microtiter plate coated with a monoclonal anti-adiponectin antibody, 2) washing of unbound materials from samples, 3) binding of a second biotinylated monoclonal anti-adiponectin antibody to the captured molecules , 4) washing of unbound materials from samples, 5) binding of streptavidin-horseradish peroxidase conjugate to the immobilized biotinylated antibody, 6) washing of excess free enzyme conjugates, and 7) quantification of immobilized antibodyenzyme conjugates by monitoring horseradish peroxidase activities in the presence of the substrate 3,3`,5,5`- tetramethylbenzidine. The enzyme activity is measured spectrophotometrically by the increased absorbance at 450 nm – 590 nm after acidification of formed products.

Since the increase in absorbance is directly proportional to the amount of captured Rat Adiponectin in the unknown sample, the latter can be derived by interpolation from a reference curve generated in the same assay with reference standards of known concentrations of Rat Adiponectin.