Overview

  • Product name

    Anti-Thrombopoietin antibody
    See all Thrombopoietin primary antibodies
  • Description

    Rabbit polyclonal to Thrombopoietin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human Thrombopoietin aa 300 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab105988)

  • Positive control

    • This antibody gave a positive signal in Human liver tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab93909 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
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 41 kDa (predicted molecular weight: 38 kDa).

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

  • Anti-Thrombopoietin antibody (ab93909) at 1 µg/ml + Human liver tissue lysate - total protein (ab29889) at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 38 kDa
    Observed band size: 41 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute
  • ICC/IF image of ab93909 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab93909 at 10µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

ab93909 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Sample
Human Cultured Cells (embryonic stem cells)
Specification
embryonic stem cells
Permeabilization
No
Fixative
Methanol

Abcam user community

Verified customer

Submitted Jun 07 2013

Answer

Thank you for contacting us.

Your credit note ID for the refund of both antibodies is xxx.

I am sorry that both antibodies did not perform as stated on their datasheets. 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

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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. 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 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.

Read More

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.

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

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Answer

Thank you for your reply.

Only ab93909 has been tested with cultured cells (ICC/IF), while ab128487 has not yet been tested in this application. I can sendyou retroactively a testing discount code for trying it.

For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein/kit on an untested species/application. You would purchase the antibody/protein/kit at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody/protein at the full price (100%) of the antibody/protein you have tested, or a full price discount for the amount paid for the kit. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.
This programapplies to ELISA kits and other kits we offer.

Please let me know.

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