Overview

  • Product name

    Human Adiponectin ELISA Kit
    See all Adiponectin kits
  • Detection method

    Colorimetric
  • Sample type

    Cell culture supernatant, Serum, Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 25 pg/ml
  • Recovery

    < 96 %

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 96.59 83% - 107%
    Serum 93.57 83% - 104%
    Plasma 90.29 80% - 103%

  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Abcam’s Human Adiponectin ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human Adiponectin in serum, plasma, and cell culture supernatants.

    This assay employs an antibody specific for Human Adiponectin coated on a 96-well plate. Standards and samples are pipetted into the wells and Adiponectin present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human Adiponectin antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Adiponectin bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Notes

    Optimisation may be required with urine samples.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    5X Assay Diluent B 1 x 15ml
    80x HRP-Streptavidin Concentrate 1 x 200µl
    Adiponectin Microplate with anti-human Adiponectin. 1 unit
    Assay Diluent A 2 x 30ml
    Assay Diluent C 2 x 15ml
    Biotinylated Anti-Human Adiponectin 2 vials
    Standard Recombinant Human Adiponectin (Lyophilized). 2 vials
    Stop Solution 1 x 8ml
    TMB One-Step Substrate Reagent 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
    • Acrp 30
    • 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
    • GBP 28
    • GBP28
    • Gelatin binding protein
    • Gelatin binding protein 28
    • Gelatin-binding protein
    • gelatin-binding protein 28
    • OTTHUMP00000210047
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab99968 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

  • Adiponectin measured in biological fluids showing quantity (pg) per mL of tested sample. Human serum and plasma were diluted 2700-24300 fold. Rat and mouse serum were diluted 1-3 fold. Human urine and saliva were diluted 300-2700 fold.

  • Representative Standard Curve using ab99968

  • Representative Standard Curve using ab99968

Protocols

References

This product has been referenced in:

  • West MD  et al. Clonal derivation of white and brown adipocyte progenitor cell lines from human pluripotent stem cells. Stem Cell Res Ther 10:7 (2019). Read more (PubMed: 30616682) »
  • Zhao L  et al. Chitosan oligosaccharide improves the therapeutic efficacy of sitagliptin for the therapy of Chinese elderly patients with type 2 diabetes mellitus. Ther Clin Risk Manag 13:739-750 (2017). Read more (PubMed: 28721055) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Abreviews
We tested this kit with rhesus macaque serum/plasma using human serum as a positive control. This kit did cross react with rhesus macaque serum/plasma, and is suitable for detecting non human primate adiponectin.

Mr. Joshua Preston

Verified customer

Submitted Mar 08 2019

Human Adiponectin ELISA Kit testing for rhesus macaques

Excellent Excellent 5/5 (Ease of Use)
Abreviews
This kit is suitable for rhesus macaques. Plasma of 17 rhesus macaques were tested and yielded concentrations comparable to human levels and other published studies of rhesus macaques.

Abcam user community

Verified customer

Submitted Nov 02 2015

Answer

Thank you for contacting us. I sent your inquiry to the lab, they did not send original data but sent the reply below. Please let me know if you have any more questions.



. A 30,000-fold dilution is quite high but our Human IgG ELISA kit actually has a recommended 10,000,000-fold dilution if you can believe that. The reason is simply that these proteins are very high in abundance in blood. Based on our QC data, it looks like when we measured normal serum and plasma that the levels of Adiponectin ranged from 2 to 120 ug/ml.



So what this means is not only is Adiponectin normally at very high levels in serum and plasma, but that it also has a large natural variance as well. Therefore, the customer should only use the recommended 30,000-fold dilution as a starting point and the optimal sample dilution would need to be determined by each customer empirically.

Read More

Answer

Thank you for contacting Abcam.

Unfortunately we have not tested pig samples with theab99968 Human Adiponectin ELISA kit, so we do not know the cross reactivity.Both antibodies used in this kit are mouse monoclonal and the epitope recognized is aa 104-244 which is in the globular domain. I did an alignement between the immunogen sequence and pig sequence, and there is about a 84% level of homology. Therefore there is a good chance that it will work for you, but we cannot guarantee it.

You could take advatage of our testing discount program, details of which are below:.

To our knowledge, ab99968 has not been tested in pig samples. Therefore, I can offer a discount off a future purchase if you buy ab99968 now, test it in pig samples and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of:$470.00 VALUE OFF OF NEXT ORDER.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab99968 in pig samples. I will then send a discount code. This code must be issued before purchasing ab99968 so please wait for my reply before ordering.

2. Purchase ab99968 either by phone, fax, or online (www.abcam.com).

3. Test it in pig samples.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any $470.00 VALUE OFF OF NEXT ORDERordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab99968 turns out to be unsuitable for pig samples, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for your enquiry.

It is possible that high viscosity of the sample can cause interference with the results. However, I would like to state that we have not tested media with such a high fat content and viscosity, therefore we do not know how it would affect the assay.

Adiponectin is normally at very high levels in serum and plasma which may resemble with the media that you are going to test. In that case, I would advise you to try perform a serial dilutions to determine the optimal dilution for a particular sample. While protease inhibitors can always be added to samples, usually they are not necessary for media samples. But if a cell or tissue lysates is going to be tested, adding protease inhibitors would be recommended.

Read More

Question
Answer

I'm happy to help! Please let me know if you need anything further.

Read More

Answer

The biotinylated anti-human adiponectin antibody is monoclonal and is raised from mice.

I hope this is helpful. Please contact us again if you have any further questions.

Read More

Answer

Thank you for your reply and for your patience. It has taken a day or two to confirm this information for you.

ab99968 Adiponectin Human ELISA Kitas been successfully tested with EDTA-plasma, heparin-plasma, and citrate-plasma.

Other anti-coagulants have not been tested, so we could notguarantee the results, but should not necessarily be excluded.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for your recent telephone enquiry.

I can confirm that the matched pair of antibodies for the Human Adiponectin ELISA kit (ab99968) have an epitope in the globular region (aa 104-244). Therefore, this kit will recognize trimers and other multimer complexes of this protein and thus detects total adiponectin (low and high molecular forms).

From QC testing of normal plasma samples with this kit, levels around 8 mg/ml were measured. This seems to be consistent with most of the reported literature available. I am sorry that as this antibody was tested externally, we do not have the raw data to provide on this occasion.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for contacting us. The capture antibody for the Adiponectin Human ELISA Kits are a monoclonal mouse antibody for kit ab99968 and a polyclonal antibody for kit ab108786 I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

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