Description
This Human Adiponectin (ACRP30) ELISA kit is used for the non-radioactive quantification of Human Adiponectin in serum, plasma, and adipocyte extracts or cell culture media samples. This kit specifically measures native Human Adiponectin and has no cross reactivity to Mouse Adiponectin. One kit is sufficient to measure 38 unknown samples in duplicate.This kit is for Research Use Only. Not for Use in Diagnostic Procedures.
This assay is a Sandwich ELISA based, sequentially, on:
1) concurrent capture of Human Adiponectin molecules from samples to the wells of a microtiter plate coated with a monoclonal anti-human adiponectin antibodies, and binding of a second biotinylated monoclonal anti-human antibody to the captured molecules ,
2) washing of unbound materials from samples,
3) binding of streptavidin-horseradish peroxidase conjugate to the immobilized biotinylated antibodies,
4) washing of excess of free enzyme conjugates, and
5) quantification of immobilized antibody-enzyme conjugates by monitoring horseradish peroxidase activities in the presence of the substrate 3,3Õ,5,5Õ-tetramethylbenzidine.
The enzyme activity is measured spectrophotometrically by the increased absorbance at 450 nm – 590nm after acidification of formed products. Since the increase in absorbance is directly proportional to the amount of captured Human 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 Human Adiponectin.
