Anti-Hemoglobin subunit alpha antibody [EP3607] (ab109444)

Overview

  • Product name

    Anti-Hemoglobin subunit alpha antibody [EP3607]
    See all Hemoglobin subunit alpha primary antibodies
  • Description

    Rabbit monoclonal [EP3607] to Hemoglobin subunit alpha
  • Host species

    Rabbit
  • Specificity

    Mouse and rat tissues tested (lung and kidney) gave negative or low sensitivity staining in IHC-P. This antibody may not be suitable for IHC with mouse or rat samples. If you have any questions, please contact our Scientific Support team.

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Hemoglobin subunit alpha aa 1-100. The exact sequence is proprietary.

  • Positive control

    • WB: K562,fetal liver, and fetal heart lysates IHC-P: Human spleen and Human colon tissues
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EP3607
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab109444 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 1/1000 - 1/10000. Detects a band of approximately 14 kDa (predicted molecular weight: 15 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Antigen retrieval is recommended.

  • Application notes
    Is unsuitable for Flow Cyt,ICC or IP.
  • Target

    • Function

      Involved in oxygen transport from the lung to the various peripheral tissues.
    • Tissue specificity

      Red blood cells.
    • Involvement in disease

      Heinz body anemias
      Alpha-thalassemia
      Alpha(0)-thalassemia is associated with non-immune hydrops fetalis, a generalized edema of the fetus with fluid accumulation in the body cavities due to non-immune causes. Non-immune hydrops fetalis is not a diagnosis in itself but a symptom, a feature of many genetic disorders, and the end-stage of a wide variety of disorders.
      Hemoglobin H disease
    • Sequence similarities

      Belongs to the globin family.
    • Post-translational
      modifications

      The initiator Met is not cleaved in variant Thionville and is acetylated.
    • Information by UniProt
    • Database links

    • Alternative names

      • Alpha 1 globin antibody
      • Alpha globin antibody
      • Alpha one globin antibody
      • Alpha-globin antibody
      • HBA_HUMAN antibody
      • HBA1 antibody
      • HBA2 antibody
      • Hemoglobin alpha 1 antibody
      • Hemoglobin alpha 1 chain antibody
      • Hemoglobin alpha 1 globin chain antibody
      • Hemoglobin alpha 2 antibody
      • Hemoglobin alpha chain antibody
      • Hemoglobin subunit alpha antibody
      • MGC126895 antibody
      • MGC126897 antibody
      see all

    Images

    • All lanes : Anti-Hemoglobin subunit alpha antibody [EP3607] (ab109444) at 1/1000 dilution

      Lane 1 : K562 lysate
      Lane 2 : Fetal liver lysate
      Lane 3 : Fetal heart lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 15 kDa
      Observed band size: 14 kDa
      why is the actual band size different from the predicted?

    • Immunohistochemical analysis of Hemoglobin subunit alpha in paraffin embedded human spleen tissue, using ab109444 at a 1/250 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of Hemoglobin subunit alpha in paraffin embedded human colon tissue, using ab109444 at a 1/250 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    References

    ab109444 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-8 of 8 Abreviews or Q&A

    Question
    Answer

    Vielen Dank für Ihren Anruf.

    Es tut mir leid, dass Sie Probleme mit diesem Antikörper hatten. Wie am Telefon besprochen habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 123456und sollteMorgen bei Ihnen ankommen.

    Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Read More

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab92492. The estimated delivery date is xxx as the item is currently backordered. I hope this is OK.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Answer

    Thank you for your reply and for testing the antibodies.

    Unfortunately, we do not have another lot in stock for ab109444 - just the same lot. Please let me know if one of the other Anti-Hemoglobin subunit alpha antibodies we carry, ab92492 or ab102758, would be a suitable replacement.

    Also, please let me know the species of the protein you tested (human, mouse or rat).
    Originally, when you had called, the antibodies gave you high background according to our records, but from the dot blot you sent it looks now more like a weak signal/no staining.
    Could you please let me know what samples were used for the original WB and what primary antibody dilution was used then? Also, please confirm if indeed high background was the issue.

    We would need this information to update our records as we evaluatethe complaints we receive.

    I look forward to hear back from you and resolve this case for you.

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

    Read More

    Answer

    According to our records, ab6721 or ab109444 was proving difficult to use in WBand we were in contact in order to help resolve the issue.

    Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced.

    As we discussed over the phone, trying a no-primary control (with secondary antibody only) would be another way to check if the problem comes from the primary or secondary antibody.

    Please let me know if thisproblem hasbeenresolved. If not, please let me know as well, so that we can resolve it in a different way (e.g. replacement, credit or refund).

    If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

    If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

    Iwish you the best of luck with your research and look forward to a reply.

    Read More

    Answer

    Thank you for contacting us.
    I am sorry to hear that one of these antibodies is notworking properly.

    As we discussed over the phone, trying a no-primary control (with secondary antibody only) would be another way to check if the problem comes from the primary or secondary antibody.

    I look forard to hear back from and assist you in resolving this problem.

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

    Read More

    Answer

    Thank you for the reply.

    The recommended dilutions on our datasheets provide a concentration to begin with, however the optimal dilution does have to be determined by the end user. Therefore I strongly recommend that you try various concentrations of the primary antibody, as there is still a good chance that this antibody will work for you, if you can identify the optimal dilution.

    Also, I understand that you are worried about high background but could you try and increase the concentration of the protein loaded to 10ug.

    If after you have tried the above suggestions you are still not seeing a signal from ab109444, then I would be happy to refund or replace the antibody.

    Please let me know if there is anything else I can help you with.

    Read More

    Question
    Answer

    Thank you for sending that protocol information.

    If I have read the information correctly, then there are a few protocol variations that may work:

    1 - Did the customer only try a 1/5000 primary antibody dilution? If so, I strongly suggest that they try range of dilutions from 1/500-1/2000, to see if they can find the concentration that will work for them.

    2 - According to the protocol, they loaded 5ug of protein on the gel. Did they try increasing the amount of protein, to say 20ug?

    3 - How long did they incubate the primary antibody for? We recommend overnight at 4C.

    4 - Did they try using 5% milk as a blocking reagent instead of BSA?

    If the customer has not tried the protocol variations above, then I would strongly suggest that they do. If they have already tried those variations, then I would be happy to process a refund for this antibody, as it is not working as we guarantee.

    I look forward to your reply.

    Read More

    Answer

    I am sorry that the replacement antibody did not prove to succesful. Unfortunately the protocol that you attached did not come through. Could you resend the protocol so that I can look it over and see if there are any suggestions that I can make.

    I look forward to your reply.

    Read More

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