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Quantikine® ELISA Kit for Measuring High Molecular Weight Adiponectin
R&D Systems now offers a Quantikine ELISA Kit for measuring high molecular weight (HMW) Adiponectin/Acrp30 in serum or plasma (Catalog # DHWAD0). Adiponectin, also known as adipocyte complement-related protein of 30 kDa (Acrp30), is a hormone secreted primarily by adipocytes that regulates numerous metabolic processes. It circulates in plasma as low molecular weight (LMW) trimers, medium molecular weight (MMW) hexamers, and HMW multimers. HMW Adiponectin is the primary bioactive form with insulin-sensitizing activity. Its circulating levels negatively correlate with obesity, insulin resistance, and coronary artery disease. The Human HMW Adiponectin/Acrp30 Kit utilizes antibodies that have been rigorously tested to ensure specificity for the HMW isoform. This is the first commercially available ELISA designed to measure HMW Adiponectin without the need for proteolytic pretreatment of the sample.
Assay Principle of the Quantikine Human HMW Adiponectin/Acrp30 ELISA Kit. A 96-well microplate pre-coated with a HMW Adiponectin-specific capture antibody is included in the Quantikine Human HMW Adiponectin/Acrp30 ELISA kit (Catalog # DHWAD0). Experimental samples or standards are added to the wells and HMW Adiponectin binds to the immobilized capture antibodies (Step 1). An HRP-conjugated HMW Adiponectin-specific detection antibody, which binds to a different epitope on HMW Adiponectin, is subsequently used to detect the bound proteins (Step 2). A tetramethylbenzidine (TMB) substrate solution is added to the wells and a blue color develops in proportion to the amount of HMW Adiponectin present in the sample. Color development is stopped, turning the color of the solution in the wells to yellow. The absorbance of the color at 450 nm is measured, producing a signal that is proportional to the amount of bound HMW Adiponectin (Step 3).
Measurement of the Levels of Human HMW Adiponectin. The mean levels of HMW Adiponectin in serum from individuals with low BMI (average BMI = 19.8; N=4), high BMI (average BMI = 32.8; N=4), and very high BMI (average BMI = 55.4; N=3) were analyzed using the Quantikine Human HMW Adiponectin/Acrp30 ELISA Kit (Catalog # DHWAD0).