ADIPONECTIN (TOTAL) ELISA KIT
INTENDED USE
The Adiponectin (Total) ELISA Kit is intended for the quantitative determination of human adiponectin in serum and plasma. It is for in vitro diagnostic use only.
INTRODUCTION
Adiponectin consists of 244 amino acids and has an approximate molecular weight of 30 kDa. The adiponectin concentration in blood is 5-30 μg/ml that accounts for about 0.01% of the total plasma proteins. Adiponectin has four domains: a signal peptide, a variable domain, a collagen-like N-terminal domain and a globular C-terminal domain. Adiponectin exists in different oligomer forms in vivo. Beside the trimer and ditrimer also high molecular multimers are present. Adiponectin is mainly expressed by adipocytes, but also muscle cells and hepatocytes can synthesize it. The only, until now known, natural inductor of the adiponectin synthesis is IGF-I. Two different adiponectin receptors are known. Both of them are ubiquitary expressed, though their distribution in the tissues varies: Adiponectin Receptor 1 (AdipoR1) is synthesized in muscle and AdipoR2 in liver tissue. The adiponectin significance for the human organism is not completely clear until now. Studies show, that adiponectin correlates negatively with BMI and could participate at the energy metabolism through the regulation of fatty acid oxidation. Adiponectin influences further physiological processes like angiogenesis.
It is also associated with glucose and lipid metabolism. Adiponectin levels are associated with insulin resistance and accordingly linked with Type II Diabetes. Furthermore, it is involved in inflammatory processes and therewith of importance for development of arteriosclerosis and coronary artery diseases. Blüher et al. (2007) showed a correlation of total serum adiponectin to insulin sensitivity and the ability of total serum adiponectin levels to predict insulin resistance and impaired glucose tolerance. Low adiponectin concentrations result in inhibition of fatty acid oxidation and are associated with insulin resistance and metabolic syndrome as well as arteriosclerosis.
PRINCIPLE OF THE TEST
Adiponectin (Total) ELISA Kit
The Adiponectin (Total) ELISA Kit is intended for the quantitative determination of human adiponectin in serum and plasma. The Adiponectin (Total) ELISA Kit assay utilizes the two-site “sandwich” technique with one monoclonal and one polyclonal antibody that bind to human Adiponectin.
Standards, controls and diluted patient samples which are assayed for human Adiponectin are added to wells of microplate coated with a high affine monoclonal anti-human Adiponectin antibody. During the first incubation step, Adiponectin in the samples is bound by the immobilized antibody. Then a peroxidase labeled conjugate is added to each well and the following complex is formed: capture antibody - human Adiponectin – Peroxidase conjugate. Tetramethylbenzidine (TMB) is used as a substrate for peroxidase. Finally, an acidic stop solution is added to terminate the reaction. The color changes from blue to yellow. The intensity of the yellow color is directly proportional to the Adiponectin concentration of sample. A dose response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated, using the values obtained from standard. Adiponectin present in the patient samples, is determined directly from this curve.
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