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  1. Link

    hepatitis-b-virus-core-antigen-antibody-c1-ab8637.pdf

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Microbiology Organism Virus DNA Virus double stranded DNA Virus Hepatitis B
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Anti-Hepatitis B Virus Core Antigen antibody [C1] (ab8637)

  • Datasheet
  • SDS
Reviews (1)Q&A (10)References (14)

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Western blot - Anti-Hepatitis B Virus Core Antigen antibody [C1] (ab8637)

    Key features and details

    • Mouse monoclonal [C1] to Hepatitis B Virus Core Antigen
    • Suitable for: ELISA, IHC-Fr, IP, WB, ICC/IF, IHC-P
    • Reacts with: Hepatitis B virus, Recombinant fragment
    • Isotype: IgG2a

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    Protein
    Recombinant Hepatitis B Virus Core Antigen protein (ab49013)

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    Overview

    • Product name

      Anti-Hepatitis B Virus Core Antigen antibody [C1]
      See all Hepatitis B Virus Core Antigen primary antibodies
    • Description

      Mouse monoclonal [C1] to Hepatitis B Virus Core Antigen
    • Host species

      Mouse
    • Specificity

      This antibody reacts with HBV Core Antigen (Major antigenic determinant, c1). Ab8637 should recognize both the precoreprotein and core protein. It will not recognize the precorprotein under native conditions, because this protein can not self assemble into particles.

      Ab8637 was raised against serotype ayw but will work with all other genotypes. 

       

    • Tested applications

      Suitable for: ELISA, IHC-Fr, IP, WB, ICC/IF, IHC-Pmore details
    • Species reactivity

      Reacts with: Hepatitis B virus, Recombinant fragment
    • Immunogen

      Tissue, cells or virus corresponding to Hepatitis B virus Hepatitis B Virus Core Antigen. Purified Hepatitis B Core Antigen

    • Epitope

      Around aa positions 70-80, major, or "c1" determinant
    • General notes

      This product was raised against native assembled HBV core (HBV capsid, or particulated core).

      For denaturing western blots and denaturing Immunofluorescence we would recommend one of the following:

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

      Anti-Hepatitis B Virus Core Antigen antibody [14E11] (ab8638)

      These products are raised to the same, but denatured, HBcAg protein sequence thus have more robust activity. 

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
    • Storage buffer

      pH: 7.4
      Constituents: 0.1% BSA, 99% PBS
    • Concentration information loading...
    • Purity

      Tissue culture supernatant
    • Clonality

      Monoclonal
    • Clone number

      C1
    • Myeloma

      Sp2/0
    • Isotype

      IgG2a
    • Light chain type

      kappa
    • Research areas

      • Microbiology
      • Organism
      • Virus
      • DNA Virus
      • double stranded DNA Virus
      • Hepatitis B
      • Cancer
      • Oncoproteins/suppressors
      • Viral proteins
      • Hepatitis B

    Associated products

    • Compatible Secondaries

      • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
      • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Isotype control

      • Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)
    • Recombinant Protein

      • Recombinant Hepatitis B Virus Core Antigen protein (ab49013)

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab8637 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
    ELISA
    Use at an assay dependent concentration.
    IHC-Fr
    1/100.
    IP
    Use at an assay dependent concentration.
    WB
    1/1000.

    We recommend boiling (100°C) the sample for 10 minutes immeditaley before loading and using a 2x or 4x Laemmli sample buffer (contains SDS). 

    A 2X concentrated Laemmli buffer is: 4% SDS, 10% beta-mercaeptoethanol, 20% glycerol, 0.1M Tris pH 6.8, and 0.005% of bromophenol blue.

    ICC/IF (1)
    Use at an assay dependent concentration.
    IHC-P
    Use at an assay dependent concentration.

    We recommend fixation for 20 minutes and incubation with primary antibody for 1 hour.

    Notes
    ELISA
    Use at an assay dependent concentration.
    IHC-Fr
    1/100.
    IP
    Use at an assay dependent concentration.
    WB
    1/1000.

    We recommend boiling (100°C) the sample for 10 minutes immeditaley before loading and using a 2x or 4x Laemmli sample buffer (contains SDS). 

    A 2X concentrated Laemmli buffer is: 4% SDS, 10% beta-mercaeptoethanol, 20% glycerol, 0.1M Tris pH 6.8, and 0.005% of bromophenol blue.

    ICC/IF
    Use at an assay dependent concentration.
    IHC-P
    Use at an assay dependent concentration.

    We recommend fixation for 20 minutes and incubation with primary antibody for 1 hour.

    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

      • Entrez Gene: 2828293 Hepatitis B virus
      • Entrez Gene: 944568 Hepatitis B virus
      • SwissProt: P03146 Hepatitis B virus
      • SwissProt: P03147 Hepatitis B virus
      • SwissProt: P03148 Hepatitis B virus
      • SwissProt: P0C573 Hepatitis B virus
      • SwissProt: P69706 Hepatitis B virus
      • 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

      Images

      • Western blot - Anti-Hepatitis B Virus Core Antigen antibody [C1] (ab8637)
        Western blot - Anti-Hepatitis B Virus Core Antigen antibody [C1] (ab8637)

        Immunoblotting analysis of the recombinant HBcAg antigen using ab8637. E.coli lysates were separated using 12% SDS-PAAG electrophoresis, imunoblotted and developed with the Amersham ECL Detection Kit. Samples: 1. Negative control; 2. HBcAg(full length, 1-183

        Immunoblotting analysis of the recombinant HBcAg antigen using ab8637. E.coli lysates were separated using 12% SDS-PAAG electrophoresis, imunoblotted and developed with the Amersham ECL Detection Kit. Samples: 1. Negative control; 2. HBcAg(full length, 1-183

      Protocols

      • Immunoprecipitation protocols
      • Immunohistochemistry protocols
      • Immunocytochemistry & immunofluorescence protocols
      • Western blot protocols

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (14)

      Publishing research using ab8637? Please let us know so that we can cite the reference in this datasheet.

      ab8637 has been referenced in 14 publications.

      • Lin Y  et al. Glucosamine promotes hepatitis B virus replication through its dual effects in suppressing autophagic degradation and inhibiting MTORC1 signaling. Autophagy 16:548-561 (2020). PubMed: 31204557
      • Xie M  et al. Scavenger receptor A impairs interferon response to HBV infection by limiting TRAF3 ubiquitination through recruiting OTUB1. FEBS J 287:310-324 (2020). PubMed: 31386800
      • Tran BM  et al. The Hepatitis B Virus Pre-Core Protein p22 Activates Wnt Signaling. Cancers (Basel) 12:N/A (2020). PubMed: 32486480
      • Lau KCK  et al. Differences in HBV Replication, APOBEC3 Family Expression, and Inflammatory Cytokine Levels Between Wild-Type HBV and Pre-core (G1896A) or Basal Core Promoter (A1762T/G1764A) Mutants. Front Microbiol 11:1653 (2020). PubMed: 32760388
      • Morrish E  et al. Combinatorial Treatment of Birinapant and Zosuquidar Enhances Effective Control of HBV Replication In Vivo. Viruses 12:N/A (2020). PubMed: 32824616
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-10 of 11 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Hepatitis B Virus Core Antigen antibody [C1]

      Good
      Abreviews
      Abreviews
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (Huh7 Human Hepatoma cells)
      Specification
      Huh7 Human Hepatoma cells
      Fixative
      Paraformaldehyde
      Permeabilization
      Yes - 0.1% Saponin
      Read More

      Abcam user community

      Verified customer

      Submitted Nov 23 2010

      Question

      The antibody ab8637 in the following order doesn't work well. The detailed informed is included in the attached file. Could you please give us some help?

      Read More

      Abcam community

      Verified customer

      Asked on Aug 20 2012

      Answer

      Thank you for contacting Abcam regarding ab8637.


      I am sorry that you are experiencing difficulties with this antibody in WB. In order to assist you further, I wanted to clarify several points in your protocol.


      You mention that you load 15ul of protein - do you know what the quantity of protein is in ug? Was the protein isolated from a particular cell type (tranfected)or translated in vitro (purified protein)? What is the difference between each of the three samples?


      Here are the recommended WB protocol conditions:


      Following lysis, prepare samples in Laemmli buffer and boil 10 minutes prior to loading.
      Load 50ng of purified protein or 50 - 100ug cell lysates
      Prior to WB, confirm efficient transfer to the membranes by Ponceau S

      WB conditions:
      Block - 5% milk (or preferrably BSA) in TBSTfor 1 hr at RT
      Primary antibody - dilute ab8637 1:1000 - 1:5000 in 5% milk (or BSA) and incubate o/n at 4oC or 1 hr at room temperature
      Wash 3X 10 min in TBST
      Secondary antibody (HRP conjugated)- dilute according to manufacturers instructions and incubate 1 hr at RT
      Wash 1X 15 min and 3X 10 min in TBST

      Develop using ECL



      Based on the results you provided, you are getting the band of interest with some background that can be cleaned up by modifying the protocol a bit. I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

      Read More

      Abcam Scientific Support

      Answered on Aug 20 2012

      Question

      Thank you for your reply.
      here is our explain for your questions:
      1. Apparently, a decreased trend was observed in the expression of the
      target protein of interest in the lanes from left to right, and the result
      from the second bloting was similar. As a matter of fact, the samples
      loaded into the 3th to the 6th lane were treated with an agent under study
      with different concentration, and an efficacy as expected was observed but
      the resultant band size was significantly larger than the expected (21KD).
      2. A SDS-PAGE gel with a percentage of 12% was used for running, and the
      samples were boiled for 10 minutes before loading.
      we are looking for your reply again

      Read More

      Abcam community

      Verified customer

      Asked on Mar 06 2012

      Answer

      Thank you for your reply.

      Upon further investigation, this target contains several phosphorylation sites which may account for the shift in molecular weight. Otherwise, I cannot account for the shift.

      If you would like to try another vial of this antibody, I would be happy to send you one. Please let me know if you are interested.

      I look forward to your reply so that I may assist you further. Please do not hesitate to contact us if you have any additional questions.

      Read More

      Abcam Scientific Support

      Answered on Mar 06 2012

      Question

      DESCRIPTION OF THE PROBLEM Non-specific band Wrong band size at the site of 35kD. SAMPLE Cell extract from HepG2.2.15. PRIMARY ANTIBODY A dilution of 1:800 was used for probing with an incubation of overnight at 4℃, then washed in TBST buffer with agitation for 4 times, 10 minutes per wash. DETECTION METHOD Pierce ECL Western Blotting Substrate Visualized by Image reader (Las 3000, Fuji film). POSITIVE AND NEGATIVE CONTROLS USED No ANTIBODY STORAGE CONDITIONS -20℃. SAMPLE PREPARATION 1% NP-40 cell lysis buffer (1% NP-40, 1 mM EDTA, 50 mM NaCl, 10 mM Tris-HCl, pH 8.0). AMOUNT OF PROTEIN LOADED 30μg per well. ELECTROPHORESIS/GEL CONDITIONS Wet transfer using nitrocellulose membrane (watman) in a standard buffer with 1X Tris-glycine omiting SDS but additioning methanol to a final concentration of 20%, as discribed by Abcam Ltd. TRANSFER AND BLOCKING CONDITIONS For membrane blocking, a 5% non-fat milk solution diluted in TBST was used. SECONDARY ANTIBODY HRP-conjugated secondary antibody against mouse IgG (, diluted in TBST with a dilution of 1:5000) was used, incubated 1h at RT and washed as same as the procedure used for primary antibody. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

      Read More

      Abcam community

      Verified customer

      Asked on Dec 21 2011

      Answer

      Thank you for contacting Abcam regarding ab8637. I am sorry that you have been experiencing difficulties with this antibody in WB. I have reviewed the protocol information and image you provided and would like to ask some additional questions. There seems to be a decrease in the expression of your protein of interest in the lanes from left to right. Were these treated differently in any way? GAPDH appears consistent, but I am not sure if you have treated the cells. What percentage gel did you run? Was everything run under reduced, denatured conditions? How long did you boil the samples before loading? It is recommended to boil the samples for at least 10 minutes for this antibody to work well. Also depending on the percentage gel and sample preparation, the molecular weight may run slightly different than the predicted molecular weight. I hope these guidelines are helpful. I look forward to your reply so that I may assist you further. Please do not hesitate to contact us if you have any additional questions.

      Read More

      Abcam Scientific Support

      Answered on Dec 21 2011

      Question

      Another question is whether Ab8637 could not distinguish HBeAg and HBcAg in cells or tissues expressing HBV whole genome. I could not totally understand these two sentences "Ab8637 should recognize both the precoreprotein and core protein.It will not recognize the precorprotein under native conditions, because this protein can not self assemble into particles." in the protocol. Thank you.

      Read More

      Abcam community

      Verified customer

      Asked on Sep 04 2007

      Answer

      Thank you for your question. I can confirm that this Hepatitis B Virus Core Antigen antibody, ab8637, will recognize native core particles but not native e antigen. Under denaturing conditions, it willr ecognize all core polypeptides, containing epitope around aa 80 (core, precore, e antigen etc.). Differentiation of core and e antigens in liver tissues using this antibody, and several others, has been published in: J Med Virol. 1997 Oct;53(2):127-38. Differentiation of core gene products of the hepatitis B virus in infected liver tissue using monoclonal antibodies.Naoumov NV, Antonov KA, Miska S, Bichko V, Williams R, Will H. This antibody will recognize all core polypeptides (core, precore, e antigen etc.) under standard western blot conditions. It appears that at least partial renaturing of proteins occurs on the membrane, so no additional renaturing steps are necessary. I hope this information is helpful. Should you have any further questions, please do not hesiate to contact us again.

      Read More

      Abcam Scientific Support

      Answered on Sep 04 2007

      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.

      Read More

      Abcam community

      Verified customer

      Asked on Sep 07 2005

      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,

      Read More

      Abcam Scientific Support

      Answered on Sep 09 2005

      Question

      thanks for the information. Is there any information that you have regarding recommended starting concentrations for use of these antibodies as either capture or detection antibodies?

      Read More

      Abcam community

      Verified customer

      Asked on Sep 07 2004

      Answer

      Here is the information that I could obtain regarding ab8639, ab8638, and ab8637. The recommended starting dilutions are 1:1,000 and down in 10-fold increments. As for detection, any known detection method would work. Once again, those mabs had being characterized in great detail in the following paper: Bichko et al. Epitopes recognized by antibodies to denatured core protein of hepatitis B virus. Mol Immunol. 1993 Feb;30(3):221-31 If you have any more questions, just let me know.

      Read More

      Abcam Scientific Support

      Answered on Sep 14 2004

      Question

      thanks for the information. Is there any information that you have regarding recommended starting concentrations for use of these antibodies as either capture or detection antibodies?

      Read More

      Abcam community

      Verified customer

      Asked on Sep 07 2004

      Answer

      I have received the following information regarding ab8255 and ab2045, but will have to get back to you about the other antibodies. The originator has said that they tested the antibodies as pairs in internal ELISA. Possible pairs are H6F5 (ab2045) - H3A4 (ab8255), and H3A4 - H6F5. They have used coating concentration 5 - 10 ug/ml and conjugate concentration 0.2 -1 ug/ml. However, the concentrations need to be optimized separately for every application.

      Read More

      Abcam Scientific Support

      Answered on Sep 13 2004

      Question

      Customer would like more details about how these antibodies were tested for application in ELISA.

      Read More

      Abcam community

      Verified customer

      Asked on Aug 26 2004

      Answer

      Thank you for your enquiry. I was able to obtain the following information regarding the use of these antibodies in ELISA. Ab8255 and Ab2045: These antibodies have been tested in ELISA with the native antigen. The antibodies can be used as a pair in sandwich ELISA. Both clones can work as capture and as detection antibodies. Ab8638, Ab8637, and Ab8639: These antibodies work well with a recombinant or natural HBcAg adsorbed on a plate, or in sandwich ELISA. Also, they work well with the synthetic HBcAg peptides. Below is one reference: Bichko et al. Epitopes recognized by antibodies to denatured core protein of hepatitis B virus. Mol Immunol. 1993 Feb;30(3):221-31

      Read More

      Abcam Scientific Support

      Answered on Sep 07 2004

      Question

      Thanks for your rapid reply. I have one more question. My end user wants to know the type of HBV core antigen.(i.e.ayw1, ayw2...) He is supposed to it's type is ayw. Inform me about that, if you can confirm it. Best Regareds

      Read More

      Abcam community

      Verified customer

      Asked on Sep 07 2004

      Answer

      Thank you for your reply. The only data we have regarding this is that it is an ayw variant as described in: Bichko et. al. Subtype ayw variant of hepatitis B virus. DNA primary structure analysis. FEBS Lett. 1985 Jun 3;185(1):208-12.

      Read More

      Abcam Scientific Support

      Answered on Sep 07 2004

      1-10 of 11 Abreviews or Q&A

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