Overview

  • Product name

    Anti-Thrombopoietin antibody [1B11]
    See all Thrombopoietin primary antibodies
  • Description

    Mouse monoclonal [1B11] to Thrombopoietin
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ELISA, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Purified recombinant fragment of Human Thrombopoietin expressed in E. coli.

  • Positive control

    • Human Thrombopoietin (amino acids 250-303) recombinant protein; MCF7 cells

Properties

Applications

Our Abpromise guarantee covers the use of ab128487 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 1/500 - 1/2000. Predicted molecular weight: 37 kDa.
ELISA 1/10000.
Flow Cyt 1/200 - 1/400.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Lineage-specific cytokine affecting the proliferation and maturation of megakaryocytes from their committed progenitor cells. It acts at a late stage of megakaryocyte development. It may be the major physiological regulator of circulating platelets.
  • Involvement in disease

    Defects in THPO are a cause of essential thrombocythemia (ET) [MIM:187950]. ET is inherited as an autosomal dominant trait which is characterized by elevated platelet levels due to sustained proliferation of megakaryocytes, and frequently lead to thrombotic and haemorrhagic complications.
  • Sequence similarities

    Belongs to the EPO/TPO family.
  • Domain

    Two-domain structure with an erythropoietin-like N-terminal and a Ser/Pro/Thr-rich C-terminal.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Database links

  • Alternative names

    • C mpl ligand antibody
    • C-mpl ligand antibody
    • Megakaryocyte colony stimulating factor antibody
    • Megakaryocyte colony-stimulating factor antibody
    • Megakaryocyte growth and development factor antibody
    • Megakaryocyte stimulating factor antibody
    • MGC163194 antibody
    • MGDF antibody
    • MKCSF antibody
    • ML antibody
    • MPL ligand antibody
    • MPLLG antibody
    • Myeloproliferative leukemia virus oncogene ligand antibody
    • Prepro thrombopoietin antibody
    • THCYT1 antibody
    • THPO antibody
    • Thrombopoietin antibody
    • Thrombopoietin nirs variant 1 antibody
    • TPO antibody
    • TPO_HUMAN antibody
    see all

Images

  • ab128487, at a 1/200 dilution, staining Thrombopoietin in MCF7 cells by Flow Cytometry (green). Negative control is shown in red.
  • ELISA using ab128487.
  • Anti-Thrombopoietin antibody [1B11] (ab128487) at 1/500 dilution + Human Thrombopoietin (amino acids 250-303) recombinant protein

    Predicted band size: 37 kDa



    Expected MWt of recombinant protein is 30 kDa.

References

ab128487 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

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.

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Answer

Thank you for your reply. I would still recommend checking if the antibodies bind to the recombinant protein in a dot blot assay. Because thrombopoietin is a secreted protein, it is very possible that it may be washed away from your cells during the staining process. If you are not satisfied with these antibodies, I will be more than happy to offer you a refund or a credit. Please let me know how you would like to proceed and I will be happy to help you further.

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Answer

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.

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Answer

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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