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    hepatitis-b-virus-x-antigen-antibody-3f6-g10-ab235.pdf

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Microbiology Interspecies Interaction Host Virus Interaction
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Anti-Hepatitis B Virus X antigen antibody [3F6-G10] (ab235)

  • Datasheet
  • SDS
Reviews (1)Q&A (4)References (10)

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Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Key features and details

  • Mouse monoclonal [3F6-G10] to Hepatitis B Virus X antigen
  • Suitable for: ELISA, IHC-P, IHC-Fr
  • Reacts with: Hepatitis B virus
  • Isotype: IgG2a

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Primary
Anti-Hepatitis B Virus Core Antigen antibody, prediluted (ab124640)
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Recombinant Enhanced GFP protein (His tag) (ab134853)
Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)

View more associated products

Overview

  • Product name

    Anti-Hepatitis B Virus X antigen antibody [3F6-G10]
    See all Hepatitis B Virus X antigen primary antibodies
  • Description

    Mouse monoclonal [3F6-G10] to Hepatitis B Virus X antigen
  • Host species

    Mouse
  • Specificity

    HB-X antigen (17 kD). We have received mixed reports regarding how ab235 works in Western blot, and we are not able to guarantee this antibody in WB.
  • Tested applications

    Suitable for: ELISA, IHC-P, IHC-Frmore details
  • Species reactivity

    Reacts with: Hepatitis B virus
  • Immunogen

    HB-Xag-Protein A Fusion protein

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

    Preservative: 0.09% Sodium azide
    Constituent: 0.1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Purified IgG prepared by affinity chromotography on protein G from tissue culture supernatant.
  • Clonality

    Monoclonal
  • Clone number

    3F6-G10
  • Myeloma

    unknown
  • Isotype

    IgG2a
  • Light chain type

    unknown
  • Research areas

    • Microbiology
    • Interspecies Interaction
    • Host Virus Interaction
    • Microbiology
    • Organism
    • Virus
    • DNA Virus
    • double stranded DNA Virus
    • Hepatitis B

Associated products

  • Compatible Secondaries

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

    • Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
  • Isotype control

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab235 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-P
1/100. This antibody may not require protein digestion pre-treatment or antigen retrieval on paraffin sections.
IHC-Fr
1/100.
Notes
ELISA
Use at an assay dependent concentration.
IHC-P
1/100. This antibody may not require protein digestion pre-treatment or antigen retrieval on paraffin sections.
IHC-Fr
1/100.

Target

  • Relevance

    Hepatitis B virus X protein (HBx) is a 17 kD transcriptional coactivator that plays a significant role in the regulation of genes involved in inflammation and cell survival. It regulates many transcription factors including nuclear factor kappa B (NF-kappaB) and plays a key role in hepatocarcinogenesis. HBx facilitates the binding of cAMP response element binding protein (CREB) to its responsive element. HBx stabilizes the cellular coactivator ASC-2 through direct protein-protein interaction, affecting the regulation of genes actively transcribed in liver cancer cells. HBx transactivates both JNK and MAPK signal transduction pathways in association with the mobilization of cytosolic Ca2+. The communication between HBx and general transcription factor TFIIB is also one of the mechanisms which account for its transcriptional transactivation. HBx decreased the expression of PTEN a known tumor suppressor and a negative regulator of phosphatidylinositol 3'-kinase/AKT and HBx decreased the expression of PTEN in HBx-transfected cells. The etiology of hepatocellular carcinoma (HCC) is involved with hepatitis B virus (HBV) infection and HBx in particular plays a role in the development of HBV-related HCC. The persistence of HBx is important to the pathogenesis of early HCC and HBx expression in the liver during chronic HBV infection may be an important prognostic marker for the development of HCC.
  • Database links

    • SwissProt: O91531 Hepatitis B virus
    • SwissProt: Q4R1S1 Hepatitis B virus
    • SwissProt: Q91C38 Hepatitis B virus
    • Alternative names

      • HBV X antigen antibody
      • HBx antibody
      • HBxAg antibody

    Protocols

    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (10)

    Publishing research using ab235? Please let us know so that we can cite the reference in this datasheet.

    ab235 has been referenced in 10 publications.

    • Song C  et al. Associations Between Hepatitis B Virus Infection and Risk of All Cancer Types. JAMA Netw Open 2:e195718 (2019). PubMed: 31199446
    • Hou Z & Quan J Hepatitis B virus X protein increases microRNA-21 expression and accelerates the development of hepatoma via the phosphatase and tensin homolog/phosphoinositide 3-kinase/protein kinase B signaling pathway. Mol Med Rep 15:3285-3291 (2017). PubMed: 28339072
    • Zhang Y  et al. Hepatitis B virus whole-X and X protein play distinct roles in HBV-related hepatocellular carcinoma progression. J Exp Clin Cancer Res 35:87 (2016). PubMed: 27255760
    • Li M  et al. Hepatitis B virus X protein promotes renal epithelial-mesenchymal transition in human renal proximal tubule epithelial cells through the activation of NF-?B. Int J Mol Med 38:513-20 (2016). PubMed: 27314843
    • Liu L  et al. Hepatitis B virus X protein mutant upregulates CENP-A expression in hepatoma cells. Oncol Rep 27:168-73 (2012). PubMed: 21956590
    • Yen CJ  et al. Hepatitis B Virus X Protein Upregulates mTOR Signaling through IKKß to Increase Cell Proliferation and VEGF Production in Hepatocellular Carcinoma. PLoS One 7:e41931 (2012). PubMed: 22848663
    • Kuo CY  et al. Apoptosis induced by hepatitis B virus X protein in a CCL13-HBx stable cell line. Oncol Rep 28:127-32 (2012). PubMed: 22552576
    • Fernandez AF  et al. The dynamic DNA methylomes of double-stranded DNA viruses associated with human cancer. Genome Res 19:438-51 (2009). WB . PubMed: 19208682
    • Kuo CY  et al. HBx inhibits the growth of CCL13-HBX-stable cells via the GSK-3beta/beta-catenin cascade. Intervirology 51:130-6 (2008). WB, IHC-P ; Human . PubMed: 18552505
    • Pál J  et al. Immunohistochemical assessment and prognostic value of hepatitis B virus X protein in chronic hepatitis and primary hepatocellular carcinomas using anti-HBxAg monoclonal antibody. Pathol Oncol Res 7:178-84 (2001). WB, IHC-P, ELISA ; Human . PubMed: 11692143

    Customer reviews and Q&As

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

    1-5 of 5 Abreviews or Q&A

    Question

    Dear Sir or Madam,

    We are a biophysical lab and we are working with recombinantly expressed HBV proteins. To validate some of our recombinant protein we're now looking into co-immunoprecipitation and western blotting. These techniques have never been used in our lab, so I have a few questions. To establish antibody techniques in our lab I am interested in HBx antibodies, but once protocols have been established we'd be interested in antibodies against other HBV proteins as well.

    Could you send me a quote for the antibodies listed below?


    Anti-6X His tag® antibody [3H2201] (ab15149)

    Anti-6X His tag® antibody (ab125262)

    Anti-6X His tag® antibody (Agarose) (ab90663)

    Anti-Hepatitis B Virus X antigen antibody [3F6-G10] (ab235)
    Anti-Hepatitis B Virus X antigen antibody [X36C] (ab2741)
    Anti-Hepatitis B Virus X antigen antibody (ab39716)

    Anti-p53 antibody [PAb 240] (ab26)

    I'm also interested in secondary antibodies for the use in western blots. Could you recommend a matching secondary anti-mouse antibody for WB use?
    WouldRabbit anti-Mouse IgG H&L (HRP) secondary antibody (ab7075) be a good choice?

    Reading the posts in your support section I can see that the epitopes of the HBx antibodies are not known. Has anything changed since then?
    And my last question would be if you supply any conformation specific antibodies against HBV core protein (HBc).

    Best wishes

    Read More

    Abcam community

    Verified customer

    Asked on Sep 03 2012

    Answer

    Sorry for the delay in getting back to you. I now have further information to share with you.

    I have checked into whether anything is know about the epitope of the anti-Hepatitis B Virus X antigen antibodies ab235, ab2741 and ab39716. I can only provide further information for ab235 additionally to the information provided on the datasheet of the antibodies. The epitope recognised by ab235 is thought to recognise an epitope of the following sequence:

    DVLCLRPVGASESR.

    I was incorrect in my last email to say that ab235 had not been tested in Western blotting. It have been tested but we have had mixed reports from customers so we would not recommend buying this antibody with that application in mind.

    I have also looked into the anti-Hepatitis B Virus Core Antigen antibodies we have and if anything is known of the conformation of the epitope is known. Unfortunately, I could not find any antibodies where this information had been established and most are able to recognise both the denatured and native form of the protein.

    I hope this information has been of help. If you have any further questions, please do not hesitate to ask.

    Read More

    Abcam Scientific Support

    Answered on Sep 03 2012

    Question

    Dear Sir or Madam,

    We are a biophysical lab and we are working with recombinantly expressed HBV proteins. To validate some of our recombinant protein we're now looking into co-immunoprecipitation and western blotting. These techniques have never been used in our lab, so I have a few questions. To establish antibody techniques in our lab I am interested in HBx antibodies, but once protocols have been established we'd be interested in antibodies against other HBV proteins as well.

    Could you send me a quote for the antibodies listed below?


    Anti-6X His tag® antibody [3H2201] (ab15149)

    Anti-6X His tag® antibody (ab125262)

    Anti-6X His tag® antibody (Agarose) (ab90663)

    Anti-Hepatitis B Virus X antigen antibody [3F6-G10] (ab235)
    Anti-Hepatitis B Virus X antigen antibody [X36C] (ab2741)
    Anti-Hepatitis B Virus X antigen antibody (ab39716)

    Anti-p53 antibody [PAb 240] (ab26)

    I'm also interested in secondary antibodies for the use in western blots. Could you recommend a matching secondary anti-mouse antibody for WB use?
    WouldRabbit anti-Mouse IgG H&L (HRP) secondary antibody (ab7075) be a good choice?

    Reading the posts in your support section I can see that the epitopes of the HBx antibodies are not known. Has anything changed since then?
    And my last question would be if you supply any conformation specific antibodies against HBV core protein (HBc).

    Best wishes

    Read More

    Abcam community

    Verified customer

    Asked on Aug 28 2012

    Answer

    Thank you for contacting us and your interest in our products.

    The request for a quote for the antibodies mentioned has been passed on to one of my colleagues who will get back to you shortly with this information. In regards to your other questions:

    The Rabbit anti-Mouse IgG H&L (HRP) secondary antibody (ab7075) should work well with ab15149, ab2741, ab235 and ab26. You would need a different secondary antibody for all the other antibodies mentioned as they are raised in rabbit. You may wantto consider the Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080). Please bear in mind that we have as yet not tested the anti-Hepatitis B Virus X antigen antibody [3F6-G10] (ab235) in Western blotting or immunoprecipitation. This does not mean that it would not work, simply that we have not tested it ourselves as yet and would therefore not he able to guarantee its performance in these applications.

    We have some good resources in setting up both immunoprecipitation and Western blotting experiments from the following link:

    https://docs.abcam.com/pdf/misc/abcam-protocols-book-2010.pdf

    Some further troubleshooting tips can also be found here:

    https://www.abcam.com/index.html?pageconfig=resource&rid=11382

    If you would like any help with this please do let us know.

    I will check if any further information is know about the epitopes recognised by the anti-HBx antibodies mentioned and will get back to you with this information as soon as I have it. I will also look into your final question ifwe have any conformation specific antibodies against HBV core protein (HBc).

    As you will be setting up these experiments from scratch, you may be interested in looking at our Western blotting range:

    https://www.abcam.com/index.html?pageconfig=resource&rid=14709&source=pagetrap&viapagetrap=optiblot

    The link above will take you not only to our Optiblot range of gels which provide superior durability and ease of use compared to many standard gels, but also the PVDF and nitrocellulose membranes and Ecl Detection kits we have available.

    If you would like any further information on these products please do let us know.

    Read More

    Abcam Scientific Support

    Answered on Aug 28 2012

    Question

    Is there a source for the recombinant protein that is mentioned on the datasheet? "Due to the viral nature of the antigen, routine antibody testing is done only on recombinant protein in Western blot."

    Read More

    Abcam community

    Verified customer

    Asked on Oct 25 2011

    Answer

    Thank you for contacting us. The note on the datasheet is from the originator of the 3F6-G10 clone. Production of the clone and testing against the recombinant protein is described in the following paper: Pál J et al. Immunohistochemical assessment and prognostic value of hepatitis B virus X protein in chronic hepatitis and primary hepatocellular carcinomas using anti-HBxAg monoclonal antibody. Pathol Oncol Res 7:178-84 (2001). PubMed: 11692143. The protein is not commercially available, as far as we know. If you are having difficulty with the antibody, please let us know and we will help resolve the problem.

    Read More

    Abcam Scientific Support

    Answered on Oct 25 2011

    Western blot abreview for Anti-Hepatitis B Virus X antigen antibody [3F6-G10]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (huh-7 cells)
    Loading amount
    20 µg
    Specification
    huh-7 cells
    Treatment
    Cells are expressing recombinant HBX protein
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 22 2009

    Question

    Our customer some questions with antibodies ab2741, ab235, ab39716. He want to know which subtype virus that these antibodis can detect? Does these antibodis detect nature protein or denatue protein?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 26 2007

    Answer

    Thank you for your message. Unfortuantley, I am unable to obtain precise information about which subtypes of Hepatitis B virus these antibodies detect. We aim to provide as much information as possible to customers, and I am sorry this is not possible on this occasion. With regards to whether the antibodies detect native or denatured protein, you should always use reducing and denaturing conditions unless stated otherwise on the datasheet. Please also check the information I have provided below for futher details. I did manage to obtain the following information on these antibodies, which I hope you will find helpful: Reference for ab2741. This reference uses the antibody in IP and WB procedures. M. Melegari, et al. Cloning and Characterization of a Novel Hepatitis B Virus x Binding Protein That Inhibits Viral Replication. J. Virology, 72(3): 1737-1743, 1998. http://jvi.asm.org/cgi/content/full/72/3/1737 Hepatitis B Virus X antigen antibody ab39716 was tested only with recombinant antigen in WB (reducing and nonreducing) and for immunoprecipitation: with antigen from HeLa cells transfected with HBX tagged with c-myc. Antibody ab235 reacts with both the native and denatured HBX antigens. For further details of this clone please see the following references: Pal J, Somogyi C, Szmolenszky A A, Szekeres G, Sipos J, Hegedus G, Martzinovits I, Molnar J, Nemeth P. Immunohistochemical assessment and prognostic value of hepatitis B virus X protein in chronic hepatitis and primary hepatocellular carcinomas using anti-HBxAg monoclonal antibody. Pathol Oncol Res. 2001;7(3):178-84. PMID: 11692143 Marczinovits I, Somogyi C, Patthy A, Nemeth P, Molnar J. An alternative purification protocol for producing hepatitis B virus X antigen on a preparative scale in Escherichia coli. J Biotechnol. 1997 Aug 11;56(2):81-8. PMID: 9304871 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 26 2007

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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