Anti-Thrombopoietin抗体(ab40664)

Thank you for your reply. If I am understanding your assay correctly, your cells are overexpressing a thrombopoietin receptor and you are incubating the cell culture with recombinant thrombopoietin to measure the binding efficacy. What type of recombinant thrombopoietin are you using? It would be very helpful if you could let me know the manufacturer and the catalog number. Have you tried a dot blot or WB with the recombinant protein to see if it is recognized by these antibodies? If you are able to detect the recombinant protein by itsself, there may be an issue with the receptor. I hope this helps, please let me know if you need any additional information or assistance.

Thank you for your reply and the additional information.

Regarding the refund, you would need to let your purchasing department know the information I had sent and they then should contact us so that the refund can be issued to you. I'm certain there is a way this can be arranged after your institution gets in touch with us.

Regarding the other 2 antibodies:
Your protocol in general looked fine and the antibody dilution used are more concentrated than the suggested range.

You mentioned that you used Hela cells and I am wondering - since 3 anti-thrombopoietin antibodies seem not to be working - if this protein is expressed in Hela cells.

It is mainly present in megakaryocytes and megakaryocytes-derived cells.
http://www.proteinatlas.org/ENSG00000090534
http://www.uniprot.org/uniprot/P40225

The protein was shown to be present also in MCF7 cells, liver tissue, HepG2 cells.
What other cell types have you tried, besides Hela cells? Would you be able to try any of the positive controls?

I look forward to hear back from you and help you further.

Thank you for contacting us.
I am sorry to hear that these antibodies do not work as expected.

According to the records in our system, the purchase of ab40664 has already been refunded and the case has been discussed and resolved.

As for ab93909 and ab128487 I have a few more questions, if you don't mind:

1) What is the actual problem you have encountered (e.g. no bands in WB, high background in ICC/IF etc)?
I just wanted to mention that ab93909 has not been tested in FACS and ab128487 has not been tested in ICC/IF, but a discount code was issued to you for trying the latter antibody in ICC/IF.

2) What antibody dilutions were used?

3) How much protein was loaded onto the gel for WB analysis?

4) For flow cytometry, did you try fixing and permeabilizing the cells?

5) How well are the secondary antibodies working with other primary antibodies?


I look forward to hear back from you and assist you further.

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Thank you for contacting us.

Your credit note ID for the refund is xxx.

I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice.

Thank you for your email.
It's OK if you do no have any images at hand.
You protocol looks fine, thus it seems the antibody is not working as it should - either the vial or the lot.

Please let me know your order or PO number. I'd be happy to replace, credit or refund the antibody.

I look forward to hear back from you and help you further.

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https://www.abcam.com/abreviews

Thank you for contacting us.

I'm sorry to hear the antibody is not working as expected.
I have not heard of this problem before for this antibody, in fact this is the first complaint we receive for this product.

Could you please let me know the following information:

1) What antibody dilsution and incubation tiem have you tried?
2) How were the cells fixed and permeabilized?
3) What reagent was used for blocking?
4) Is the secondary antibody working well with other primary antibodies?
5) Could you send me an image of your various sample types?

I look forward to hear back from you and assist you further.

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https://www.abcam.com/abreviews