Cat No.: 32012 Other Names: Acrp30, AdipoQ, apM1, GBP-28
Introduction
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.
Assay Performance
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.
Adiponectin (ng/mL) | Absorbance (450 nm) | Blanked absorbance |
0 | 0.056 | 0 |
7.8 | 0.105 | 0.049 |
15.6 | 0.158 | 0.102 |
31.2 | 0.255 | 0.199 |
62.5 | 0.463 | 0.407 |
125 | 0.845 | 0.789 |
250 | 1.608 | 1.552 |
500 | 2.599 | 2.543 |
B. Sensitivity
The lowest level of adiponectin that can be detected by this assay is 7.8 ng/mL.
C. Specificity
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.
D. Precision
Intra-assay Precision (Precision within an assay) C. V.<10%.
Inter-assay Precision (Precision between assays) C.V.<10%.
E. Recovery
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%.