Anti-Thyroid Peroxidase/TPO 抗体 [MoAb47] (ab12500)

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.

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.

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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.

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.

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.