Adiponectin ELISA Kit, mouse
Description:
Adiponectin ELISA Kit, mouse
Trade Name:
Chemicon (Millipore)
Product Overview:
Introduction
It is becoming clear that adipose tissue is not merely a storage place for excess energy unused but secretes a number of biologically active soluble factors collectively named adipocytokines, thereby regulating glucose and fatty acid metabolism. The gene encoding adiponectin was identified as an adipose tissue-specific gene by screening a subtractive adipocyte cDNA library (1). The gene product was independently identified as gelatin-binding protein- 28 (GBP28) (2). The mouse orthologue was isolated by several groups and designated adipocyte complement-related protein of 30kDa (Acrp30), adipocyte most abundant protein-1 (APM1), or AdipoQ (3-4). Albeit differently being named it was unequivocally concluded that adiponectin is an adipocyte-specific protein and represents a major serum protein. The full-length adiponectin in plasma exists as trimer, hexamer, and multimer whereas extremely low amount of globular domain itself also exists in plasma as trimer (5). Due to its adipocyte-specific expression it was thought that adipo-nectin could be responsible for regulation of adipocyte physiology. In many rodent models, it has been shown that infusion of adiponectin ameliorates hyperglycemia and hyperinsulinemia
(6-8). One notable mechanism for decreasing blood sugar is that adiponectin inhibits hepatic glucose production by downregulating the key enzymes involved in gluconeogenesis such as phophoenolpyruvate carboxy-kinase and glucose-6-phosphatase (9). When skeletal muscle cells or a muscle cell line is treated with adiponectin, both glucose uptake and fatty acid oxidation are significantly upregulated (6-8). These studies suggests an unifying conclusion that adiponectin regulates both systemic and hepatic insulin resistance. Direct evidence was shown that administration of adiponectin reduces atherosclerosis in apolipoof protein E-deficient mice (10). Therefore, measurement serum adiponectin levels gives us important information on the role of adiponectin in regulation of glucose and/or lipid metabolism.
Test Principle
This kit is enzyme-linked immunosorbent assay (ELISA) for quantitative determination of Adiponectin in mouse serum. Monoclonal antibody specific for mouse Adiponectin has been pre-coated onto 96-well microplate. Standards and samples are pipetted into the wells and any adiponectin present is bound by immobilized antibody. Bound adiponectin is captured by purified anti-mouse adiponectin polyclonal antibody. HRP conjugated anti-rabbit IgG is added. After washing, a substrate solution is added. The colors develop in proportion to the bounded adiponectin quantity. The color development is stopped and the intensity of color is measured.
It is becoming clear that adipose tissue is not merely a storage place for excess energy unused but secretes a number of biologically active soluble factors collectively named adipocytokines, thereby regulating glucose and fatty acid metabolism. The gene encoding adiponectin was identified as an adipose tissue-specific gene by screening a subtractive adipocyte cDNA library (1). The gene product was independently identified as gelatin-binding protein- 28 (GBP28) (2). The mouse orthologue was isolated by several groups and designated adipocyte complement-related protein of 30kDa (Acrp30), adipocyte most abundant protein-1 (APM1), or AdipoQ (3-4). Albeit differently being named it was unequivocally concluded that adiponectin is an adipocyte-specific protein and represents a major serum protein. The full-length adiponectin in plasma exists as trimer, hexamer, and multimer whereas extremely low amount of globular domain itself also exists in plasma as trimer (5). Due to its adipocyte-specific expression it was thought that adipo-nectin could be responsible for regulation of adipocyte physiology. In many rodent models, it has been shown that infusion of adiponectin ameliorates hyperglycemia and hyperinsulinemia
(6-8). One notable mechanism for decreasing blood sugar is that adiponectin inhibits hepatic glucose production by downregulating the key enzymes involved in gluconeogenesis such as phophoenolpyruvate carboxy-kinase and glucose-6-phosphatase (9). When skeletal muscle cells or a muscle cell line is treated with adiponectin, both glucose uptake and fatty acid oxidation are significantly upregulated (6-8). These studies suggests an unifying conclusion that adiponectin regulates both systemic and hepatic insulin resistance. Direct evidence was shown that administration of adiponectin reduces atherosclerosis in apolipoof protein E-deficient mice (10). Therefore, measurement serum adiponectin levels gives us important information on the role of adiponectin in regulation of glucose and/or lipid metabolism.
Test Principle
This kit is enzyme-linked immunosorbent assay (ELISA) for quantitative determination of Adiponectin in mouse serum. Monoclonal antibody specific for mouse Adiponectin has been pre-coated onto 96-well microplate. Standards and samples are pipetted into the wells and any adiponectin present is bound by immobilized antibody. Bound adiponectin is captured by purified anti-mouse adiponectin polyclonal antibody. HRP conjugated anti-rabbit IgG is added. After washing, a substrate solution is added. The colors develop in proportion to the bounded adiponectin quantity. The color development is stopped and the intensity of color is measured.
Key Applications:
ELISA
Application Notes:
The CHEMICON® Mouse Adiponectin ELISA Kit is designed for in vitro quantitative detection of adiponectin in serum samples of mouse origin. There are enough reagents included in this kit for one 96-well immunoassay plate. Running duplicate wells for samples and standards is recommended.
For Research Use Only; Not for use in diagnostic procedures
For Research Use Only; Not for use in diagnostic procedures
Species Reactivity:
Mouse
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Descriptive Text:
Sensitivity:
Sensitivity: 50 pg/mL
Intra-assay Variation: +/- 4.15 % (15.52 µg/mL)
Inter-assay Variation: +/- 7.88 % (15.95 µg/mL)
Recovery: 98~105% for spiked samples
Cross-reactivity: No cross-reactivity with human and rat sera. No reactivity detected with 10 ng/mL of human and rat adiponectin. No reactivity detected with 100 ng/mL of mouse Resistin, RELM-beta, Leptin, or human TNF-alpha.
Intra-assay Variation: +/- 4.15 % (15.52 µg/mL)
Inter-assay Variation: +/- 7.88 % (15.95 µg/mL)
Recovery: 98~105% for spiked samples
Cross-reactivity: No cross-reactivity with human and rat sera. No reactivity detected with 10 ng/mL of human and rat adiponectin. No reactivity detected with 100 ng/mL of mouse Resistin, RELM-beta, Leptin, or human TNF-alpha.
Therapeutic Areas:
Obesity
Packaging:
96 wells
Materials Required but Not Delivered:
1. Precision single and multi-channel pipettes.
2. Disposable pipette tips.
3. Microtubes or equivalent for preparing dilutions.
4. Disposable plastic containers for preparing working detector antibody
and substrate.
5. Reagent reservoirs.
6. Microwell or microstrip plate reader 450 nm.
7. Deionized water.
2. Disposable pipette tips.
3. Microtubes or equivalent for preparing dilutions.
4. Disposable plastic containers for preparing working detector antibody
and substrate.
5. Reagent reservoirs.
6. Microwell or microstrip plate reader 450 nm.
7. Deionized water.
Components:
- 96-Well Plate: - (Catalog No. CYT286a) 12x8 well strips coated with absorbed monoclonal antibody against mouse adiponectin.
- Wash Concentrate: - (Catalog No. CYT286b) One 100 mL (5X) bottle.
- Assay Diluent: - (Catalog No. CYT286c) One 50 mL (10X) bottle.
- Secondary Antibody: - (Catalog No. CYT286d) Rabbit anti-Mouse Adiponectin Polyclonal Antibody, 12 mL.
- QC Sample: - (Catalog No. CYT286e) Positive control having 16-22 µg/mL range of mouse serum adiponectin.
- Detector: - (Catalog No. CYT286f) One 150 µL (100X) bottle of HRP conjugated anti-rabbit IgG.
- Adiponectin Standard (Recombinant Mouse): - (Catalog No. CYT286g) One 16.0 ng vial (Lyophilized).
- Substrate I: - (Catalog No. CYT286h) One 6 mL bottle.
- Substrate II: - (Catalog No. CYT286i) One 6 mL bottle.
- Stop Solution: - (Catalog No. CYT286j) One 12 mL bottle.
Kit or Assay Type:
ELISA Assays
Assay Type:
ELISA Assays
Storage Conditions:
Reagents must be stored at 2° to 8°C when not in use. Reagents must be brought to room temperature before use. Do not expose reagents to temperatures greater than 25°C. Diluted wash solution may be stored at room temperature for up to one month.