Human Adiponectin ELISA Kit

Human Adiponectin ELISA Kit

SKU: 31010

 

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Assay range : 0.78-100 ng/ml

Size :      96T/Kit

 

  • 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 quantitative sandwich ELISA using monoclonal antibodies against human adiponectin. The immunoplate is pre-coated with a monoclonal antibody specific for human adiponectin. Standards and samples are pipetted into the wells and any human adiponectin present is bound by the immobilized antibody. After washing away any unbound substances, a horseradish peroxidase (HRP)-linked monoclonal antibody specific for human adiponectin is added to the wells. After a final wash step, 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

    0.068

    0

    1.56

    0.112

    0.044

    3.12

    0.156

    0.088

    6.25

    0.248

    0.18

    12.5

    0.418

    0.35

    25

    0.724

    0.656

    50

    1.384

    1.316

    100

    2.469

    2.401

     

    B. Sensitivity:

    The lowest level of human adiponectin that can be detected by this assay is 1.56 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 including human resistin, TNFa, ANGPTL4, insulin, leptin and IL6.

     

    D. Precision:

    The assay variations of this ELISA kits were studied on four human serum samples with varying concentrations of endogenous adiponectin. The mean within variation was calculated from results of five duplicate determinations in each assay of the indicated samples. The mean between variations of each sample was calculated from results of four separate assays with duplicate samples in each assay.

    Sample No.

    Mean Adiponectin Levels (mg/ml)

    Within% CV

    Between% CV

    1

    9.21

    4.02

    4.97

    2

    21.33

    3.65

    4.68

    3

    5.32

    3.27

    4.53

    4

    15.72

    4.18

    5.01

     

    E. Recovery:

    Varying amounts of human adiponectin were added to three human serum samples and the Adiponectin content was determined in three separate assays. The % of recovery = observed adiponectin concentrations/expected adiponectin concentrations x 100%.

     

    Average recovery

    Range (%)

    1

    99.3

    97-104

    2

    99.8

    98-103

    3

    101.1

    99-105

    4

    100.2

    98-104

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