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Human Total Adiponectin/Acrp30 ELISA - Quantikine Best Seller

R&D Systems, part of Bio-Techne | Catalog # DRP300

R&D Systems, part of Bio-Techne
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DRP300
PDRP300
SRP300

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)

Sensitivity

0.891 ng/mL

Assay Range

3.9-250 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Human Total Adiponectin/Acrp30 Quantikine ELISA

The Quantikine Human Total Adiponectin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure total (low, middle, and high molecular weight) human Adiponectin in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human Adiponectin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Adiponectin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring Adiponectin.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human

Specificity

Natural and recombinant human total Adiponectin (low, middle, and high molecular weight)

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human Adiponectin in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Serum (n=60)6641865 - 21,4243665
EDTA plasma (n=35)55481198 - 19,9733557
Heparin plasma (n=35)60261359 - 20,6913728

Cell Culture Supernates -  Human peripheral blood leukocytes (1 x 106 cells/mL) were cultured in DMEM supplemented with 5% fetal bovine serum, 50 μM beta-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernate were removed and assayed for levels of total human Adiponectin. No detectable levels were observed.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 12.5 45.3 91.5 12.7 47.1 100
Standard Deviation 0.41 1.27 4.56 1.00 2.80 6.70
CV% 3.2 2.8 5.0 7.9 5.9 6.7

EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (ng/mL) 19.8 69.9 143 20.5 74.4 157
Standard Deviation 0.50 2.40 6.76 1.40 4.30 10.8
CV% 2.5 3.4 4.7 6.8 5.8 6.9

Recovery for Human Total Adiponectin/Acrp30 Quantikine ELISA

The recovery of Adiponectin spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 101 88-112

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Adiponectin were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human Total Adiponectin ELISA/Acrp30 ELISA Linearity

Scientific Data Images for Human Total Adiponectin/Acrp30 Quantikine ELISA

Human Total Adiponectin ELISA/Acrp30 ELISA Cell Culture Supernate Standard Curve

Human Total Adiponectin ELISA/Acrp30 ELISA Cell Culture Supernate Standard Curve

Human Total Adiponectin ELISA/Acrp30 ELISA Serum/Plasma Standard Curve

Human Total Adiponectin ELISA/Acrp30 ELISA Serum/Plasma Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Adiponectin/Acrp30

Adiponectin, alternately named Adipocyte complement-related protein of 30 kDa (Acrp30), adipoQ, adipose most abundant gene transcript 1 (apM1), and gelatin-binding protein of 28 kDa (GBP28), is an adipocyte-specific, secreted protein with potential roles in glucose and lipid homeostasis. Circulating Adiponectin levels are high, accounting for approximately 0.01% of total plasma protein (1-4). Adiponectin contains a modular structure that includes an N-terminal collagen-like domain followed by a C-terminal globular domain with significant sequence and structural resemblance to the complement factor C1q (1, 5, 6). Although they share little sequence identity, similar threedimensional structure and certain conserved amino acid residues suggest an evolutionary link between the C1q-like domain of Adiponectin and members of the TNF superfamily (7). Adiponectin assembles into different complexes including trimers (low molecular weight), hexamers (middle molecular weight), and higher order oligomeric structures (high molecular weight) that may affect biological activity (1, 7, 8). Adiponectin is induced during adipocyte differentiation and its secretion is stimulated by insulin (1, 9). Two receptors for Adiponectin, termed AdipoR1 and AdipoR2, have been cloned (10). Although functionally distinct from G-protein-coupled receptors, the genes encode predicted proteins containing 7 transmembrane domains. AdipoR1 is highly expressed in skeletal muscle, while AdipoR2 is primarily found in hepatic tissues. 
Injection of Adiponectin into non-obese diabetic mice leads to an insulin-independent decrease in glucose levels (11). This is likely due to insulin-sensitizing effects involving Adiponectin regulation of triglyceride metabolism (11). A truncated form of Adiponectin (gAdiponectin) containing only the C-terminal globular domain has been identified in the blood, and recombinant gAdiponectin has been shown to regulate weight reduction as well as free fatty acid oxidation in mouse muscle and liver (2, 12). The full-length recombinant Adiponectin protein is apparently less potent at mediating these effects (2, 12). The mechanism underlying the role of Adiponectin in lipid oxidation may involve the regulation of expression or activity of proteins associated with triglyceride metabolism including CD36, acyl CoA oxidase, AMPK, and PPAR gamma (12-14). 
Although Adiponectin-regulation of glucose and lipid metabolism in humans is less clear, similar mechanisms may also be in place (15). A negative correlation between obesity and circulating Adiponectin has been well established (6, 16, 17), and Adiponectin levels increase concomitantly with weight loss (18). Decreased Adiponectin levels are associated with insulin resistance and hyperinsulinemia, and patients with type-2 diabetes are reported to exhibit decreased circulating Adiponectin (19, 20). Thiazolidinediones, a class of insulin-sensitizing, anti-diabetic drugs, elevate Adiponectin in insulin-resistant patients (21). In addition, high Adiponectin levels are associated with a reduced risk of type-2 diabetes (22). Using magnetic resonance spectroscopy it has been demonstrated that intracellular lipid content in human muscle negatively correlates with Adiponectin levels, potentially due to Adiponectin-induced fatty acid oxidation (15). 
Adiponectin may also play anti-atherogenic and anti-inflammatory roles. Adiponectin plasma levels are decreased in patients with coronary artery disease (20). Furthermore, neointimal thickening of damaged arteries is exacerbated in Adiponectin-deficient mice and is inhibited by exogenous Adiponectin (23). Adiponectin inhibits endothelial cell expression of adhesion molecules in vitro, suppressing the attachment of monocytes (24). In addition, Adiponectin negatively regulates myelomonocytic progenitor cell growth and TNF-alpha production in macrophages (25, 26).

Alternate Names

Acrp30, AdipoQ, ApM1, GBP28

Entrez Gene IDs

9370 (Human); 11450 (Mouse); 246253 (Rat)

Gene Symbol

ADIPOQ

Additional Adiponectin/Acrp30 Products

Product Documents for Human Total Adiponectin/Acrp30 Quantikine ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Total Adiponectin/Acrp30 Quantikine ELISA

For research use only

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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.