Overview

  • Product name
  • Description
    Rabbit polyclonal to Adiponectin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein (E. coli).

  • Positive control
    • Human Serum.

Properties

Applications

Our Abpromise guarantee covers the use of ab13881 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
WB
  • Application notes
    WB: 1/1000. Predicted molecular weight: 30 kDa.

    Not tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • 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
    • Database links
    • Alternative names
      • 30 kDa adipocyte complement related protein antibody
      • 30 kDa adipocyte complement-related protein antibody
      • ACDC antibody
      • ACRP30 antibody
      • ADIPO_HUMAN antibody
      • Adipocyte antibody
      • Adipocyte C1q and collagen domain containing protein antibody
      • Adipocyte complement related 30 kDa protein antibody
      • Adipocyte complement related protein of 30 kDa antibody
      • Adipocyte complement-related 30 kDa protein antibody
      • adipocyte-specific secretory protein antibody
      • Adiponectin antibody
      • Adiponectin precursor antibody
      • adiponectin, C1Q and collagen domain containing antibody
      • Adipoq antibody
      • Adipose most abundant gene transcript 1 antibody
      • Adipose most abundant gene transcript 1 protein antibody
      • Adipose specific collagen like factor antibody
      • ADIPQTL1 antibody
      • ADPN antibody
      • APM 1 antibody
      • apM-1 antibody
      • APM1 antibody
      • C1q and collagen domain-containing protein antibody
      • GBP28 antibody
      • Gelatin binding protein antibody
      • Gelatin binding protein 28 antibody
      • Gelatin-binding protein antibody
      see all

    Images

    • SDS-PAGE 12% separation of human adiponectin 5µg / line.

      SDS-PAGE 12% separation of human adiponectin 5µg / line.

    References

    This product has been referenced in:
    • Ma Z  et al. Epigenetic drift of H3K27me3 in aging links glycolysis to healthy longevity in Drosophila. Elife 7:N/A (2018). Read more (PubMed: 29809154) »
    • Rocha N  et al. Human biallelic MFN2 mutations induce mitochondrial dysfunction, upper body adipose hyperplasia, and suppression of leptin expression. Elife 6:N/A (2017). WB ; Human . Read more (PubMed: 28414270) »
    See all 4 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Answer

    Dear Dr Steinhoff, (I sent this e-mail to you last week, but today received an automated reply from your mail server suggesting that it did not reach you, so I forward again just in case) Thank you for sending us the mass spec data for this antibody. I've reviewed your correspondence with Charlotte and chatted to her about your feedback. I've also confirmed that there are no protein stabilisers included with this antibody e.g. BSA that could be contributing to the additional peaks. Another idea is that the additional peaks could be corresponding to degraded IgG molecules - I'd be happy to send you another vial from a different batch for you to test. We sell the antibody for use in Western Blotting (WB), and as already advised do not recommend it for use in mass spec or any other procedures requiring a pure preparation of IgGs. Whole serum is often used in WB where all sorts of other proteins will be present alongside the IgG. That's why the blocking and washing steps are so important to eliminate the non-specific binding of other molecules that are present alongside the IgG of interest. Although the antibody has been affinity purified with the immunogen, it may well be that other proteins have not all been eliminated. If so, I agree with your comment that the protocol for immunogen affinity purification needs to be optimised. Let me know if you would like me to send you another vial for you to test. Also, if you'd like to discuss this matter over the telephone, I can be reached via +44 1223 472283 (direct line). Best regards

    Read More

    Question

    Thank you very much for your email. However, I have to admit that I strongly disagree with your reply: 1. "Ab13881 is a polyclonal product as such multiple isoforms of IgG will recognise different epitopes on the immunognen that the product is purified with. The different isoforms of IgG may be expected to give different mass spectra." The structure of all IgG molecules is such that they consist of four polypeptide chains: two heavy chains (of approximately 50kDa) and two light chains (of about 25kDa). While there are two types of light chains (? and ?) there are five types of heavy chains which represent the different isoforms: IgM, IgD, IgG, IgA and IgE. Both heavy and light chains contain constant as well as variable domains with the variable domains being responsible for antigen binding. The only antibody region differing from one polyclonal IgG molecule to the other will be the variable region that does NOT represent the majority of amino acids and thus will not significantly effect the overall mass of the antibody. Since the according to your product insert the isotype in the solution is "IgG" this is no explanation at all for the peaks I see in the QC experiment of your antibody. 2. Ab13881 is not an ultra pure product, it is immunogen affinity purified for use in Western blotting but not for sensitive analytical protocols requiring unltra pure products. Again, I have to disagree on your comment. If an antibody is immunogen affinity purified I expect the fraction to be clean: aim of this step is to "select" for only the antibodies recognizing the antigen of interest. If I get a spectrum with multiple peaks not only in the high but also in the low mass range the protocol for the immunogen affinity purification needs to be optimized. Even if I use the antibody for Western Blot I need to be sure that I only add my antibody of interest and not other molecules that will bind and effect the outcome of my experiment! See below for a spectrum of an immunogen purified polyclonal antibody. This is what you expect to see. I have done these kind of experiments a lot and the Acrp30 antibody is the first immunogen affinity purified antibody that is not clean. 3. "Monoclonal products may be more pure but there will still be peaks expected other than the Ab dependant on the buffer constituents." Yes and no: since you state that your antibody is in a solution of 0.1M PBS, 0.1M Sodium Chloride I would also expect it to be clean. Only if there are protein impurities I will see them in the mass spec. No matter if I'm looking at an immunogen affinity purified polyclonal antibody or a monoclonal antibody the solution needs to be clean = "purified". Would you please be so kind as to send me the contact details for the sales person responsible for Switzerland? I think it will be important to discuss these issues with her/him. I think it is in the interest of Abcam to learn about the problems we encountered and to be avoid problems like this in the future. Thank you very much,

    Read More
    Answer

    Thank you for sending us the mass spec data for this antibody. I've reviewed your correspondence with Charlotte and chatted to her about your feedback. I've also confirmed that there are no protein stabilisers included with this antibody e.g. BSA that could be contributing to the additional peaks. Another idea is that the additional peaks could be corresponding to degraded IgG molecules - I'd be happy to send you another vial from a different batch for you to test. We sell the antibody for use in Western Blotting (WB), and as already advised do not recommend it for use in mass spec or any other procedures requiring a pure preparation of IgGs. Whole serum is often used in WB where all sorts of other proteins will be present alongside the IgG. That's why the blocking and washing steps are so important to eliminate the non-specific binding of other molecules that are present alongside the IgG of interest. Although the antibody has been affinity purified with the immunogen, it may well be that other proteins have not all been eliminated. If so, I agree with your comment that the protocol for immunogen affinity purification needs to be optimised. Let me know if you would like me to send you another vial for you to test. Also, if you'd like to discuss this matter over the telephone, I can be reached via +44 1223 472283 (direct line). Best regards

    Read More

    Question

    Sorry for the delay in getting back to you after my phone call last week. Please find below the mass spectrum of the QC I did with the above antibody. The way the method works is that we have an array with a "normal phase chemistry" and I applied 1ul of the above antibody that will bind to the array and is detected in our reader. For a clean antibody preparation you expect to see a peak at 140-150kDa (depending on the species) and another peak at half of the mass representing mainly the double charged intact antibody. As you can clearly see in the figure below is that there are a lot more peaks - meaning proteins - around than the intact antibody (and there are more impurities below 20kDa). The problem is that the experiments I wanted to do require a clean antibody preparation since the antibody is covalently linked by reaction with primary amines to a matrix - which will of course mean that we will not only link the antibody but all the other molecules as well, risking unspecific binding to the molecules and loosing capacity for the reaction itself. From your datasheet it sounds to me that the antibody solution should be clean - but as you can clearly see above this is not the case so that this antibody preparation is of no use for us. Would you please be so kind as to let me know if this is only a problem of the lot or if this is a general issue with this particular antibody? I don't know if you have access to a SELDI system - this QC experiment took me only 10min to perform and provides a lot of crucial information. Thanks in advance for your help!

    Read More
    Answer

    Thank you for your image. We are sorry that you have been experiencing problems with this product. Ab13881 is a polyclonal product as such multiple isoforms of IgG will recognise different epitopes on the immunognen that the product is purified with. The different isoforms of IgG may be expected to give different mass spectra. Ab13881 is not an ultra pure product, it is immunogen affinity purified for use in Western blotting but not for sensitive analytical protocols requiring unltra pure products. Monoclonal products may be more pure but there will still be peaks expected other than the Ab dependant on the buffer constituents. Best regards

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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