Anti-Hepatitis B Virus Core Antigen antibody [10E11] (ab8639)

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Overview

  • Product name
    Anti-Hepatitis B Virus Core Antigen antibody [10E11]
    See all Hepatitis B Virus Core Antigen primary antibodies
  • Description
    Mouse monoclonal [10E11] to Hepatitis B Virus Core Antigen
  • Host species
    Mouse
  • Specificity

    This antibody reacts with HBV Core Antigen (amino acid residues 1-10). Ab8639 should recognize both the precoreprotein and core protein.

    Ab8639 was raised against serotype ayw but will work with all other genotypes. 

  • Tested applications
    Suitable for: ICC/IF, WB, IP, ELISA, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Hepatitis B virus
  • Immunogen

    Purified Denatured Hepatitis B Core Antigen

  • Epitope
    aa positions 1-10

Properties

Applications

Our Abpromise guarantee covers the use of ab8639 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 1/100.
WB 1/1000.
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC-P 1/100.
IHC-Fr 1/100.

Target

  • Relevance
    Hepatitis B Virus Core Antigen (HBcAg) is part of the infectious virion containing an inner "core particle" enclosing the viral genome. The icosahedral core particle contains 180 or 240 copies of the core protein. HBcAg is one of the three major clinical antigens of hepatitis B virus but disappears early in the course of infection. The hepatitis B virus core antigen (HBcAg) is a highly immunogenic subviral particle and functions as both a T-cell-dependent and a T-cell-independent antigen. Therefore, HBcAg may be a promising candidate target for therapeutic vaccine control of chronic HBV infection.
  • Cellular localization
    Capsid protein: Virion. Host cytoplasm, hepatocyte nucleus.
  • Database links
    • Alternative names
      • C antibody
      • Capsid protein antibody
      • Core and e antigen antibody
      • core antibody
      • Core antigen antibody
      • Core protein antibody
      • HBc antibody
      • HBcAg antibody
      • HBVgp4 antibody
      • Hepatitis B core antigen antibody
      • Hepatitis B Virus core antigen antibody
      • p21.5 antibody
      • precore/core protein antibody
      see all

    References

    This product has been referenced in:
    • Liu K  et al. Multiple roles of core protein linker in hepatitis B virus replication. PLoS Pathog 14:e1007085 (2018). Read more (PubMed: 29782550) »
    • Peyret H  et al. Tandem fusion of hepatitis B core antigen allows assembly of virus-like particles in bacteria and plants with enhanced capacity to accommodate foreign proteins. PLoS One 10:e0120751 (2015). WB . Read more (PubMed: 25830365) »
    See all 4 Publications for this product

    Publishing research using ab8639? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    Submit a review Submit a question

    1-10 of 15 Abreviews or Q&A

    Question

    Thank you and I appreciate you taking some care over this.
    Obviously, if you look at our account, you will see we use a huge amount of this. A substantial change in titre would make this very expensive indeed and we would need some form of recompense to make up for the reduced amount of antibody.

    Regards

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    Answer

    Thank you very for your cooperation in this case. I apologize for being late as our collaborator lab was unable to get back to me in time.

    At Abcam we have a strict ethical policies, we care about animals and always track if our collaborator labs are fulfilling the same ethical terms and conditions. This was the reason why the product was changed from Ascites to tissue culture supernatant. Indeed, the titters of ascites were higher for this mab (as well as for many others, this is typical), but the titters of the bioreactor supernatants are also high, in full accordance with specifications.

    We acknowledge the inconvenience this may have caused which is totally due to nature of antibody production. We are happy to provide 5% discount in your next purchase or free shipping as a goodwill gesture discount. Could you please contact me for the code?

    I hope this information is never the less helpful to you. Should you have any question please do not hesitate to contact me.

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    Question

    I am not really sure that this is sorting the problem at all. Rather than just take pot luck, would it not be sensible to send us a small aliquot of whatever batches you have to test first?
    Changing the antibody is completely out of the question; the ones we have chosen are for specific parts of the protein and also work on western blots. The others do not and are therefore of no use whatsoever to us.
    I propose that you prepare a list of stock and then send us a small sample of each to test.

    Regards

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    Answer

    We unfortunately do not keep small samples of antibodies so we are unable to send you sample of each lot. The only thing we can do is to ship one vial of new lot.

    Let me know if you are interested in receivingone free of charge vial.

    Thank you very much for your cooperation.

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    Question

    I'm afraid the discount is somewhat missing the point. What we really want is for the antibody to be concentrated prior to sale. The new batch is massively less effective than the tissue culture supernatant. Obviously, XXXXXXXXX is welcome, but it does not address the fact that our bills have gone up nearly 50% as a result of Abcam selling an inferior product.

    Regards

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    Answer

    Thank you very much for your patience. I apologize for the delay as it took me longer than expected, due to interdepartmental communications.

    I have checked the previous history of this antibody and found that the purity of this antibody was actually changed 5 years back and not this year. This means you received the same antibody purity throughout i.e. in the form of tissue culture supernatant rather than Ascites. We have notsold this antibody as purified, so the low activity observed is unlikey to be due to a change in purity. This could be due to a change in protocol, batch to batch variations or one off bad vial etc. Could you confirm whether the protocol was changed between the lots? Did you observe any difference with other lots?

    It is true that we were unable to update the datasheet in time, and we apologize for this. This antibody has never been purified so we do not know the concentration dependant activity of this product. I can assure you that, the quality of this antibody was always of high standard and each lot gave good results. We were unaware of any problem before your call.

    This investigation has revealed that, nothing recently changed with this antibody; and unfortunately as a result we are unable to determine, why you have observed low activity. We appreciate your cooperation in this whole case and as a valued customer; XXXXXXXXX


    I will also give you a call after lunch in order to know more about the problem and any progress you have made in past few weeks.

    Once again many thanks for your cooperation. Speak to you soon.

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    Question

    Thank you and I appreciate you taking some care over this.
    Obviously, if you look at our account, you will see we use a huge amount of this. A substantial change in titre would make this very expensive indeed and we would need some form of recompense to make up for the reduced amount of antibody.

    Regards

    Read More
    Answer

    I am currently having discussions with lab. I will get back to you soon.

    Many thanks for having patience!

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    Question

    Thanks for this.
    The most obvious question is why is the antibody the same price when it is clearly at a significantly lower titre?

    Regards

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    Answer

    Thank you for contacting us.

    We unfortunately do not know the concentration and titre of the product because it is not a purified antibody. Generally the titre of tissue culture supernatant is lower than the ascites, which was the basis of my response. Because we hadn't determined the concentration of this product so it will be hard to guess how much difference in the titre was.

    I have contacted our lab regarding the QC data of both lots and now waiting a reply from them, I will be able to send you this info soon.

    Thank you very much for having patience. Please do not hesitate to contact us if you need any more advice or information.

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

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    Question

    please find attached completed questionnaire and collated results (in ppt file) with regard to the decreased efficiency of the anti-HBV core antigen monoclonal antibody [10E11] (product code: ab8639; lot: GR17742-1), which we have routinely bought from you with variable results. Upon investigation it is now apparent that the problem goes back to earlier experiments where we have used the same lot number. The good results have been whenever we have gone to frozen stocks of an earlier batch (lot: 637899)
    We rely upon this antibody for routine immunoassays and are therefore very keen to learn why there has been such a marked reduction in apparent titre in the recent batch.

    Regards,

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    Answer

    Thank you very much for your email and for filling the questionnaire. I am very sorry to hear about the problems you are experiencing.

    The most significant change happened between the lot #637899 and lot #GR17742-1 is the purity of the antibody. The lot #637899 was produced as ascites and the lot GR17742 was produced as tissue culture supernatant. The titre (concentration) of antibodies in ascites is always higher than the tissue culture supernatants, this may explain the difference in results.

    The datasheet of this antibody was changed in February this year; reflecting the correct information about the product (purity). I suppose this info might have missed when ordering the recent lot.

    To overcome this change we always suggest our customer changing the dilution factor. Have you tried any troubleshooting with the current lot?

    I hope this information will be useful. I will look forward to hearing from you soon for further assistance.

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    Question

    The lot 637899 works well in western blot however GR17742-1 produces very faint band.
    - dilution 1/300
    - Same protocol used between different lots

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    Answer

    Thank you for calling us and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    As we discussed on the phone, I am attachingour questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

    I look forward to receiving your reply.

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    Question

    1. Order details: * Batch number: 140202 * Abcam order or Purchase order number: ab8639 * Antibody storage conditions (temperature/reconstitution etc) Working antibody store at 4?, Aliquot and store at -80? 2. Please describe the problem (high background, wrong band size, more bands, no band etc). Wrong band size. 3. On what material are you testing the antibody in WB? · Species: human, Huh7 cell transfected with pHBV-48. · Cell extract or Nuclear extract: cell extract · Purified protein or Recombinant protein: 3. The lysate * How much protein was loaded: 30 mg * What lysis buffer was used: RIPA lysis buffer (150 mM NaCl, 50 mM Tris pH 7.5, 10 mM EDTA, 1% NP-40, 0.1% SDS); Triple-detergent lysis buffer (150 mM NaCl, 50 mM Tris pH 8.0, 1% NP-40, 0.1% SDS, 0.5% sodium deoxycholate) * What protease inhibitors were used: Aprotinin, Leupeptin, PMSF * What loading buffer was used: 4X SDS sample buffer ( 250 mM Tris-HCl pH 6.8, 8% SDS, 20% b-mercaptoethanol, 40% Glycerol, 0.04% Bromophenol Blue) * Did you heat the samples: temperature and time: 99?, 10 min 4. Electrophoresis/Gel conditions/ Transfer conditions * Reducing or non reducing gel: SDS PAGE * Gel percentage : 12% * Transfer conditions: Buffer: 25 mM Tris, 192 mM Glycine, 20% Methanol 250 mA, 2 hr 5. Blocking conditions * Buffer: PBS * Blocking agent: milk, BSA, serum, what percentage: 5% skim milk * Incubation time: 1 hr * Incubation temperature: RT 6. Primary Antibody * Specification (in which species was it raised against): human hepatitis B virus core antigen · At what dilution(s) have you tested this antibody: 1:1000 or 1:500 · What dilution buffer was used: 5% skim milk/PBS · Incubation time: 1 overnight · Incubation temperature: 4? · What washing steps were done: TBS-T (0.5% Tween-20) wash 10 min for 3 times 7. Secondary Antibody * Specification (in which species was it raised against)? mouse * At what dilution(s) have you tested this antibody: 1:5000 * Incubation time 1hr * Wash steps: TBS-T (0.5% Tween-20) wash 10 min for 3 times * Do you know whether the problems you are experiencing come from the secondary? I don’t think so. 8. Detection method ECl, ECl+, other detection method: Immobilon TM Western Detection Reagents 9. Background bands · Have you eliminated the possibility that any background bands could be due to the secondary antibody? (Run a “No primary” control): no. · Is the blocking step sufficient? Yes. · Are your washing steps sufficiently stringent? (Multiple short washes are more effective than fewer longer wash steps) Yes · At what size are the bands migrating? Could they be degradation products of your target? 21 kDa. · Please provide an image of your blot (as an e-mail attachment, a faxed image is not sufficient) Fig.1. anti-HBc (Abcam, 10E11), 1:1000 Fig. 2. anti-HBc (Abcam, 10E11), 1:500 Fig. 3. anti-HBc (homemade rabbit polyclonal antibody), 1:1000 as a control. Protein samples on fig. 1 and fig.3 are the same. Fig. 4. anti-HBs ([another company] rabbit polyclonal antibody), 1:4000 as a control. Membrane in fig.1 was stripped and reprobe with anti-HBs antibody. Did you apply positive and negative controls along with the samples? Please specify. Positive control: HBV virion, negative control: Huh7 cell lysate 10. Optimization attempts · How many times have you tried the Western? 2 times · Do you obtain the same results every time e.g. are background bands always in the same place? Not exactly the same. · What steps have you altered? I have tried different concentrations (1:1000 and 1:500) and three kinds of protein extraction method: 1) RIPA-1: RIPA lysis buffer?scape the cells?Lysis on ice for 20 min?centrifuge 13,000 rpm, 20 min? Collect the supernatant and add sample buffer to the pellet and sonicate. 2) RIPA-2: RIPA lysis buffer?scape the cells?freeze and thaw?centrifuge 13,000 rpm, 20 min? Collect the supernatant and add sample buffer to the pellet and sonicate. 3) 3-dtgn: triple-detergent lysis buffer?scape the cells?freeze and thaw?sontcation?whole lysate.

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    Answer

    Thank you for providing extensive details of the customer's protocol, as well as images, it enabled me to assess very rapidly that there was a problem with the vial of ab8639 that you received. I can offer a replacement vial or credit note, whichever the customer prefers, thank you for letting me know what he/she would like,

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    Question

    What is the immunogen used to generate ab8638,ab 8639, ab8637, it is meant to recognise the core protein of Hepatitis B, can we confirm this is the core and NOT the precore protein. The customer wants to detect the core protein of Hep B, in particular fragment 1-150 (N term) in WB.

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    Answer

    Ab8637 and ab8639 should recognize the precorerotein in WB but there is no "cross-reaction" as all proteins are distinguishable by mobility on the gel (i.e they will recognise both the precoreprotein and core protein). AB8638 may or may not recognize your protein fragment aa1-150, since its epitope is aa aa35-140 of the core protein, which is really close to the N terminus of the protein you are going to analyze. Ab8637 will not recognize the precorprotein under native conditions, because this protein can not self assemble into particles. I hope this information will help you, please let me know if you need further assistance,

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    Question

    Does this antibody recognize the HBsAg subtype adw?

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    Answer

    Thank you for your enquiry. We don not have information as to whether it recognize the HBsAg subtype adw. Ab8639 reacts with HBV Core Antigen (amino acid residues 1-10) and this antibody has been characterized in great detail in the following paper: Bichko V et al. Epitopes recognized by antibodies to denatured core protein of hepatitis B virus. Mol Immunol 30:221-31 (1993). PubMed: 7679466

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