Overview

  • Product name
    Anti-Thyroid Peroxidase/TPO antibody [MoAb47]
    See all Thyroid Peroxidase/TPO primary antibodies
  • Description
    Mouse monoclonal [MoAb47] to Thyroid Peroxidase/TPO
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Full length native protein (purified) corresponding to Human Thyroid Peroxidase/TPO.

  • Positive control
    • Thyroid

Properties

Applications

Our Abpromise guarantee covers the use of ab12500 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
IHC-P
  • Application notes
    IHC-P: 1/10 - 1/25 in an ABC method. Perform heat mediated antigen retrieval with 1mM EDTA buffer, pH 9.0 before commencing with IHC staining protocol. We suggest an incubation period of 30-60 minutes at room temperature.

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

    • Function
      Iodination and coupling of the hormonogenic tyrosines in thyroglobulin to yield the thyroid hormones T(3) and T(4).
    • Pathway
      Hormone biosynthesis; thyroid hormone biosynthesis.
    • Involvement in disease
      Note=An alternative splicing in the thyroperoxidase mRNA can cause Graves' disease.
      Defects in TPO are the cause of congenital hypothyroidism due to dyshormonogenesis type 2A (CHDH2A) [MIM:274500]; also called genetic defect in thyroid hormonogenesis 2A or thyroid hormone organification defect II. CHDH2A is due to defective conversion of accumulated iodide to organically bound iodine. The iodide organification defect can be partial or complete.
    • Sequence similarities
      Belongs to the peroxidase family. XPO subfamily.
      Contains 1 EGF-like domain.
      Contains 1 Sushi (CCP/SCR) domain.
    • Post-translational
      modifications
      Glycosylated.
      Heme is covalently bound through a H(2)O(2)-dependent autocatalytic process. Heme insertion is important for the delivery of protein at the cell surface.
      Cleaved in its N-terminal part.
    • Cellular localization
      Membrane and Cell surface.
    • Information by UniProt
    • Database links
    • Alternative names
      • MSA antibody
      • PERT_HUMAN antibody
      • TDH2A antibody
      • Thyroid microsomal antigen antibody
      • Thyroid peroxidase antibody
      • Thyroperoxidase antibody
      • TPO antibody
      • TPX antibody
      see all

    References

    This product has been referenced in:
    • Cairns DM  et al. Muscle cells enhance resistance to pro-inflammatory cytokine-induced cartilage destruction. Biochem Biophys Res Commun 392:22-8 (2010). Read more (PubMed: 20043873) »
    • Cairns DM  et al. The role of muscle cells in regulating cartilage matrix production. J Orthop Res 28:529-36 (2010). Read more (PubMed: 19813241) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    1-5 of 5 Abreviews or Q&A

    Question
    Answer

    Thank you for your enquiry.

    I can confirm that ab12500 Anti-Thyroid Peroxidase antibody [MoAb47]antibody is sold as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

    I can confirm that for tissue culture supernatant, concentration of antibody is known to very between 1 - 3 mg/ml.

    I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you.

    For peptide blocking, usually use twice the amount of peptide to antibody although please note that this may require some optimization:
    https://www.abcam.com/index.html?pageconfig=resource&rid=11378

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

    Read More

    Answer

    Thank you for contacting us.

    According to the datasheet, this antibody is a non-purified cell culture supernatant product. That means, along with whole serum andascitis products, the antibody concentration cannot be determined as there are too many other proteins in this solution as well.


    However, we have a table under our Technical FAQs (Question 8, What concentration of primary antibody should I use?) which explains how much specific antibody can be expected in the non-purified products.


    As a supernatant, the antibody concentration should be between 1 - 3 mg/ml. I would therefore suggest to use a concentration of 6 mg/ml, to be on the safe side.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Question

    My customer is using using Biotin-XX goat anti-mouse IgG (H+L) secondary antibody and it is a highly sensitive biotin-based detection method, here is the details of the product :http://products.invitrogen.com/ivgn/product/W10132 Based on the last result (dot blot) that i got, there was totally no signal at 1/500 dilution or 16ul in 8ml of washing buffer (serum based blocking buffer, incubated at room temp for 1hour) for this protein ( The signal that showed in the picture that attached in the previous email is the result of another antibody Moab47. The purpose of showing this result is to prove the method is working!!!). Apart from this, i have also tried different concentration of sample (up to 160ug!!!) but still failed to detect any signal. Since other users managed to detect the protein by using only 20 ug of total cell lysate and antibody dilution at 1:4000, i don't think i should repeat my experiment with 1/250 dilution antibody!!! For you information, it is no way for the customer to keep on trying with different concentration as the customer is wasting his reagents. The customer really need a good and workable solution so that he can solve his problem. Obviously, this is not the detection problem and also not the method problem as it is working in other antibodies. For you information, my customer antibody is only left one reaction and what he need is a good solution and method to help him solve the problem that he got. Thank you very much and hope to receive your soonest reply. Thanks again. Best regards,

    Read More
    Answer

    Thank you for providing some further information. I understand your customer has used up most of the product during testing and optimization. This is to let you know that I have confirmed and agreed on placing a new order for your customer - for one vial ofab818 (from batch: GR59271-2) as a free of charge replacement. I hope the second vial will work as it is expected, and please do let me know how you are getting on with this product. Dear Colleague, Please add 1 vial ofab818 (from a different batch: GR59271-2) to the next shipment for Bitalifsc. Original CCE ID: XXXXXXX Original Order:XXXXXXX Problem is: wrong band size on Western blot and no signal in Dot blot Many thanks.

    Read More

    Answer

    Thank you for getting back to me and for providing some further details. I understand that whilst ab818 failed to work In Western blot and in Dot blot; the other antibody ab12500 seemed to be fine. It is important to mention that these antibodies detect different target proteins (TBP and Thyroid Peroxidase, respectively) so they can't be used as positive control. The localization of these proteins are also different as well as the purity and the concentration of the antibodies (Protein G purified and Tissue culture supernatant). 1) Localisation: TBP protein is localized in the cell nuclei so it is recommended preparing nuclear lysate and using it for Western blot application. As the datasheet indicates HeLa nuclear extract can be applied as positive control. This is the best way to find out if the antibody is still active or not. 2) Dilution/concentration: Has the customer tried different dilution i.e. 1/500, 1/250 to see if the signal is getting stronger. This is particular important since whole cell lysate – rather than nuclear lysate - was applied. 3) Detection system: I understand that the secondary antibody was Biotin-XX goat anti-mouse IgG (H+L). Could you explain if the detection is ECL/ECL Plus or other type? Does it include an enzyme/fluorochrome conjugated system? In case it was not ECL system, optimization of the primary antibody would be even more essential I look forward to hearing from you soon.

    Read More

    Answer

    Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet, these are updated as soon as any new information is brought to our attention. As far as we are aware, cross reactivity with rat has not yet been tested for use with ab12500. Should you decide to go ahead and purchase this product, please let us know how you get on by submitting an Abreview and in return we will award you Abpoints, which can be redeemed on a number of rewards (a further 100 points will be offered for an image). You can find our IHC protocol at: https://www.abcam.com/assets/pdf/protocols/IHC-paraffin%20protocol%20(IHC-P).pdf This antibody is recommended to be used at 1/10 to 1/25 in an ABC method. You should perform heat mediated antigen retrieval with 1mM EDTA buffer (pH 9.0) before commencing with IHC staining protocol. We suggest incubating with the primary for 30-60 minutes at room temperature. Please let me know if you require any further assistance.

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