Overview

  • Product name
    Human Adiponectin ELISA Kit
    See all Adiponectin kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 5 3.2%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 4.3%
  • Sample type
    Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    7 pg/ml
  • Range
    39.1 pg/ml - 2500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 99 95% - 106%
    Cell culture media 86 74% - 97%
    Heparin Plasma 104 93% - 117%
    EDTA Plasma 98 94% - 103%
    Citrate Plasma 96 87% - 100%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
  • Product overview

    Adiponectin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Adiponectin protein in serum, plasma, and cell culture supernatant.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Adiponectin is a 30kDa monomer that combines to form trimers (low molecular weight), hexamers (middle molecular weight), and other higher molecular weight multimers. ab222508 is formulated to denature these complexes and measure the total Adiponectin. Adiponectin is an important adipokine involved in fat metabolism and insulin sensitivity.   Insulin stimulates the secretion of adiponectin. Decreased levels of adiponectin are associated with insulin resistance, whereas high levels of adiponectin are associated with reduced risk of type-2 diabetes.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human Adiponectin Capture Antibody 1 x 600µl
    10X Human Adiponectin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    Denaturant 1 x 500µl
    Human Adiponectin Lyophilized Recombinant Protein 1 x 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas
  • Function
    Important adipokine involved in the control of fat metabolism and insulin sensitivity, with direct anti-diabetic, anti-atherogenic and anti-inflammatory activities. Stimulates AMPK phosphorylation and activation in the liver and the skeletal muscle, enhancing glucose utilization and fatty-acid combustion. Antagonizes TNF-alpha by negatively regulating its expression in various tissues such as liver and macrophages, and also by counteracting its effects. Inhibits endothelial NF-kappa-B signaling through a cAMP-dependent pathway. May play a role in cell growth, angiogenesis and tissue remodeling by binding and sequestering various growth factors with distinct binding affinities, depending on the type of complex, LMW, MMW or HMW.
  • Tissue specificity
    Synthesized exclusively by adipocytes and secreted into plasma.
  • Involvement in disease
    Defects in ADIPOQ are the cause of adiponectin deficiency (ADPND) [MIM:612556]. ADPND results in very low concentrations of plasma adiponectin.
    Genetic variations in ADIPOQ are associated with non-insulin-dependent diabetes mellitus (NIDDM) [MIM:125853]; also known as diabetes mellitus type 2. NIDDM is characterized by an autosomal dominant mode of inheritance, onset during adulthood and insulin resistance.
  • Sequence similarities
    Contains 1 C1q domain.
    Contains 1 collagen-like domain.
  • Domain
    The C1q domain is commonly called the globular domain.
  • Post-translational
    modifications
    Hydroxylated Lys-33 was not identified in PubMed:16497731, probably due to poor representation of the N-terminal peptide in mass fingerprinting.
    HMW complexes are more extensively glycosylated than smaller oligomers. Hydroxylation and glycosylation of the lysine residues within the collagene-like domain of adiponectin seem to be critically involved in regulating the formation and/or secretion of HMW complexes and consequently contribute to the insulin-sensitizing activity of adiponectin in hepatocytes.
    O-glycosylated. Not N-glycosylated. O-linked glycans on hydroxylysines consist of Glc-Gal disaccharides bound to the oxygen atom of post-translationally added hydroxyl groups. Sialylated to varying degrees depending on tissue. Thr-22 appears to be the major site of sialylation. Higher sialylation found in SGBS adipocytes than in HEK fibroblasts. Sialylation is not required neither for heterodimerization nor for secretion. Not sialylated on the glycosylated hydroxylysines. Desialylated forms are rapidly cleared from the circulation.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Alternative names
    • 30 kDa adipocyte complement related protein
    • 30 kDa adipocyte complement-related protein
    • ACDC
    • ACRP30
    • ADIPO_HUMAN
    • Adipocyte
    • Adipocyte C1q and collagen domain containing protein
    • Adipocyte complement related 30 kDa protein
    • Adipocyte complement related protein of 30 kDa
    • Adipocyte complement-related 30 kDa protein
    • adipocyte-specific secretory protein
    • Adiponectin
    • Adiponectin precursor
    • adiponectin, C1Q and collagen domain containing
    • Adipoq
    • Adipose most abundant gene transcript 1
    • Adipose most abundant gene transcript 1 protein
    • Adipose specific collagen like factor
    • ADIPQTL1
    • ADPN
    • APM 1
    • apM-1
    • APM1
    • C1q and collagen domain-containing protein
    • GBP28
    • Gelatin binding protein
    • Gelatin binding protein 28
    • Gelatin-binding protein
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab222508 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Adiponectin were measured in duplicates, interpolated from the Adiponectin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:5,000, plasma (EDTA) 1: 2,500, plasma (heparin) 1: 5,000 and plasma (citrate) 1: 5,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Adiponectin concentration was determined to be 7962 ng/mL in serum, 6106 ng/mL in plasma (EDTA), 7377ng/mL in plasma (heparin) and 7671 ng/mL in plasma (citrate).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Adiponectin concentration was determined to be 9,188 ng/mL with a range of 3,048 – 47,443 ng/mL.

  • The concentrations of Adiponectin were measured in duplicates, interpolated from the Adiponectin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum free cell culture media 2.5% (1:40). The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Protocols

References

ab222508 has not yet been referenced specifically in any publications.

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