Intended use
This immunoassay kit allows for the specific measurement of human Adiponectin concentrations in cell culture supernates, serum, and plasma.
Introduction
Adiponectin, alternatively named Adipocyte Complement-Related Protein of 30 kDa (Acrp30), shares structural similarity with complement factor C1q and is a member of the family of defense collagens. It is secreted exclusively by differentiated adipocytes and circulates at high concentrations (inug/mL range). Adiponectin has a modular structure comprising an N-terminal collagenous domain with multiple collagen triple helix repeats, followed by a C-terminal C1q-like globular domain. The globular domain has similar folding topology with tumor necrosis factor-a and assembles into homotrimers. Higher order oligomeric adiponectins (hexamers and higher molecular weight forms) are also formed via interactions between the collagenous stalk. A truncated form of Adiponectin containing only the globular domain (gAdiponectin or gAcrp30) can be generated by proteolytic cleavage. The gAdiponectin as well as all oligomeric forms of the full length Adiponectin are detected in serum. Different isoforms of Adiponectin have been shown to activate different signal transduction pathways. Conflicting biological activities have been reported for the various isoforms.
Two seven membrane-spanning Adiponectin receptors, designated AdipoR1 and AdipoR2 have been identified. AdipoR1 is expressed predominantly in muscle and functions as a high-affinity receptor for gAdiponectin, but a very low-affinity receptor for the full length Adiponectin. AdipoR2 binds both the full length and globulin domain with intermediate affinity and is expressed primarily in liver. Adiponectin is an anti-diabetic and anti-atherogenic hormone that plays important roles in the regulation of lipid and glucose metabolism.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for Adiponectin has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any Adiponectin present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for Adiponectin is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells
and color develops in proportion to the amount of Adiponectin bound in the initial step. The color
development is stopped and the intensity of the color is measured.
