Rapid Human Adiponectin
Adiponectin is an adipokine exclusively expressed in adipose tissues with potent anti-diabetic, anti-atherogenic and anti-inflammatory functions.
In humans, decreased serum adiponectin levels are associated with increased body mass index (BMI), decreased insulin sensitivity, less favourable plasma lipid profiles, increased inflammation and increased risk for the development of type 2 diabetes, hypertension and coronary heart diseases.
PRINCIPLE OF THE ASSAY
This assay is a sandwich ELISA designed for the quantitative detection of human adiponectin in samples in 1 hour. A mouse monoclonal antibody specific to human adiponectin has been pre-coated onto a micro-titre plate. The user pipettes standards and samples into the wells and any human adiponectin present is sandwiched by the immobilised antibody and a second horseradish peroxidase (HRP)-linked monoclonal antibody specific to human adiponectin that is co-incubated with the samples. After wash step to remove any unbound reagents, an HRP substrate solution is added and colour develops in proportion to the amount of human adiponectin bound initially. The assay is stopped and the optical density of the wells determined using a microplate reader. Since the increases in absorbance are directly proportional to the amount of captured human adiponectin, the unknown sample concentration can be interpolated from a reference curve included in each assay.
A. Typical representation of standard curve
The following standard curve is provided for demonstration only. A standard curve should be generated for each set of sample assay.
Absorbance (450 nm)
The lowest level of adiponectin that can be detected by this assay is 3.9 ng/ml.
The antibody pair used in this assay is specific to human adiponectin and does not cross-react with mouse and rat adiponectin, and other cytokine or hormone molecules tested, including human resistin, TNFα, ANGPTL4, insulin, leptin and IL6.
Intra-assay Precision (Precision within an assay) C. V <10%. Inter-assay Precision (Precision between assays) C.V <10%.
The recovery of the assay was determined by adding various amounts adiponectin to a sample. The measured concentration of the spiked sample in the assay was compared to the expected concentration. The average recovery was 91%.