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Anti-HCV subtype 1b NS5B antibody (ab35586)

Reviews (1)Q&A (6)References (17)
Western blot - Anti-HCV subtype 1b NS5B antibody (ab35586)

    Key features and details

    • Rabbit polyclonal to HCV subtype 1b NS5B
    • Suitable for: ELISA, WB
    • Reacts with: Hepatitis C virus
    • Isotype: IgG

    Overview

    • Product name

      Anti-HCV subtype 1b NS5B antibody
      See all HCV subtype 1b NS5B primary antibodies

    • Description

      Rabbit polyclonal to HCV subtype 1b NS5B

    • Host species

      Rabbit

    • Tested applications

      Suitable for: ELISA, WBmore details

    • Species reactivity

      Reacts with: Hepatitis C virus

    • Immunogen

      Recombinant fragment (Hepatitis C Virus) with a 21 amino acid C-terminal truncation

    • General notes

      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

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab35586 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 (1)
    Use at an assay dependent dilution.
    WB
    1/2000. Detects a band of approximately 64 kDa.
    Notes
    ELISA
    Use at an assay dependent dilution.
    WB
    1/2000. Detects a band of approximately 64 kDa.

    Target

    • Cellular localization

      Host cytoplasm, nucleus, mitochondrion, endoplasmatic reticulum; secreted.

    • Database links

      • Alternative names

        • Hepacivirin antibody
        • NS3P antibody
        • POLG_HCVBK antibody
        • Serine protease NS3 antibody

      Images

      • Western blot - Anti-HCV subtype 1b NS5B antibody (ab35586)
        Western blot - Anti-HCV subtype 1b NS5B antibody (ab35586)
        All lanes : Anti-HCV subtype 1b NS5B antibody (ab35586) at 1/2000 dilution

        Lane 1 : Lysate from Huh7 cells, mock transfection
        Lane 2 : Lysate from Huh7 cells transfected with NS5B

        Observed band size: 64 kDa why is the actual band size different from the predicted?

      References (17)

      Publishing research using ab35586? Please let us know so that we can cite the reference in this datasheet.

      ab35586 has been referenced in 17 publications.

      • Malikova AZ  et al. Pre-Senescence Induction in Hepatoma Cells Favors Hepatitis C Virus Replication and Can Be Used in Exploring Antiviral Potential of Histone Deacetylase Inhibitors. Int J Mol Sci 22:N/A (2021). PubMed: 33925399
      • Balakrishnan S  et al. A Nondimensional Model Reveals Alterations in Nuclear Mechanics upon Hepatitis C Virus Replication. Biophys J 116:1328-1339 (2019). PubMed: 30879645
      • Zhao Q  et al. Dual Roles of Two Isoforms of Autophagy-related Gene ATG10 in HCV-Subgenomic replicon Mediated Autophagy Flux and Innate Immunity. Sci Rep 7:11250 (2017). PubMed: 28900156
      • Vegna S  et al. NOD1 Participates in the Innate Immune Response Triggered by Hepatitis C Virus Polymerase. J Virol 90:6022-35 (2016). PubMed: 27099311
      • Valero ML  et al. Hepatitis C Virus RNA-Dependent RNA Polymerase Interacts with the Akt/PKB Kinase and Induces Its Subcellular Relocalization. Antimicrob Agents Chemother 60:3540-50 (2016). PubMed: 27021315
      View all Publications for this product

      Customer reviews and Q&As

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

      1-7 of 7 Abreviews or Q&A

      ELISA abreview for Anti-HCV subtype 1b NS5B antibody

       Good
      Abreviews
      abreview image
      Application
      ELISA
      Sample
      Human Cell (huh-7 hepatocarcinoma cells)
      Specification
      huh-7 hepatocarcinoma cells
      Blocking step
      BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C
      Type
      Sandwich (Capture)
      Read More
      The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

      Abcam user community

      Verified customer

      Submitted Jun 17 2008

      Question

      Thank you for the free antibody replacement.

      We would like to take another lot of the same antibody (ab35586).

      We noticed that abcam offers two database links on the datasheet of ab35586

      http://www.uniprot.org/uniprot/P26663 Hepatitis C Virus
      http://www.uniprot.org/uniprot/O39928 Hepatitis C Virus

      We would like to know which one database link listed above is referred to the immunogen designed to produce ab35586?

      We will be appreciated that you can share more detail information (ex. immuonogen sequence) about this antibody.

      Read More
      Answer

      Thank you for contacting us.

      The lab sent me the correct immunogen info:
      "Recombinant HCV (Hepatitis C Virus) NS5B with a 21 amino acid C-terminal truncation, genotype 1b, clone Con1, UniProt link
      http://www.uniprot.org/uniprot/?query=taxonomy:333284"

      We will update the datasheet accordingly.
      Here is the information about your free of charge replacement:
      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 a new lot.
      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.
      Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

      Read More

      Question

      hier ist der ausgefüllte Fragebogen.
      Mit freundliche Grüssen,

      1) Abcam product code: ab100895
      2) Abcam order reference number or product batch number:
      3) Description of the problem: Nonspecific binding on Western blot, no specific bands with genotype 2A
      4) Sample preparation:
      Type of sample (whole cell lysates, fraction, recombinant protein…): whole cell lysate
      Lysis buffer: RIPA
      Protease inhibitors: PMSF, AEBSF, 104 mM, Aprotinin, 80 μM,Bestatin, 4 mM
      E-64, 1.4 mM, Leupeptin, 2 mM, Pepstatin A, 1.5mM
      Reducing agent:: SDS, 2-mercaptoethanol
      Boiling for ≥5 min? yes
      Protein loaded ug/lane: 70 µg
      Positive control: yes, HCV core / ns5a antibody staining
      Negative control: yes, uninfected cell lysate
      5) Percentage of gel
      Type of membrane: Hybond ECL
      Protein transfer verified: yes, marker is well transferred and other antibodies work well
      Blocking agent and concentration: 10% milk-TBST
      Blocking time: 1h
      Blocking temperature: RT
      6) Primary antibody (If more than one was used, describe in “additional notes”) : HCV NS5b, HCV Core, HCV NS5a
      Concentration or dilution 1:1000
      Diluent buffer: 1% milk-TBST
      Incubation time: over night
      Incubation temperature: 4°C
      7) Secondary antibody: goat-anti mouse HRP labeled
      Species: goat
      Reacts against: mouse
      Concentration or dilution 1:10000
      Diluent buffer: 1% milk-TBST
      Incubation time: 1h
      Incubation temperature: RT
      Fluorochrome or enzyme conjugate: HRP
      8) Washing after primary and secondary antibodies:
      Buffer: TBST
      Number of washes: 3 times
      9) Detection method: ECL
      10) How many times have you run this staining? 2x
      Do you obtain the same results every time? yes
      What steps have you altered to try and optimize the use of this antibody?
      Document attachment: Attaching images of your blot is strongly recommended and can greatly speed up our investigation of your problem.

      Read More
      Answer

      Vielen Dank dafür, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.



      Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten.

      Um Ihr Problem zu lösen, möchte ich die folgenden Vorschläge zu Veränderung Ihres Protokolls machen:

      1. Um eine Maskierung Ihres Zielproteins zu vermeiden, möchte ich vorschlagen, den Gehalt der Blocksubstanz auf maximal 5% abzusenken. Wie meine Kollegin schon anmerkte, haben wir die Erfahrung gemacht, dass BSA in vielen Faellen das bessere Blockmittel ist.

      2. Um die Nebenbanden abzuschwächen, mochte ich vorschlagen die Proteinladung auf 30 ug pro Bahn zu reduzieren: So sollte noch genügend Zielprotein vorhanden sein, allerdings wird die Möglichkeit der unspezifischen Bindungen reduziert.

      3. Leider wurden keine Angaben zum Gel gemacht: Handelt es sich um ein natives oder reduzierendes Gel? Die meisten unserer Antikörper wurden nur unter denaturierenden/ reduzierenden WB Bedingungen getestet. Ich möchte daher vorschlagen, ein reduzierendes Gel zu verwenden, falls Sie dies nicht schon tun.


      Bitte lassen Sie mich wissen, ob ich Ihnen helfen konnte. Wenn ein Antikörper nicht so funktioniert wie wir diesen auf unserem Datenblatt beschreiben, bieten wir unseren Kunden gerne einen kostenlosen Ersatz, eine Gutschrift oder eine Erstattung an, wenn das Produkt wie auf dem Datenblatt beschrieben behandelt und innerhalb von 180 Tagen nach Erhalt reklamiert wurde.

      Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.




      Read More

      Question

      Sehr geehrtes Abcam-Team,
      vielen Dank für die Antwort bezüglich des NS5b Antikörpers. Können Sie mir schon sagen, ob eine Kreuzreaktion mit dem HCV Genotyp 2a möglich ist?
      Viele Grüße

      Read More
      Answer

      Vielen Dank dafür, dass Sie nachhaken.

      Leider haben wir noch keine guten Nachrichten für Sie:

      Wie versprochen haben wir uns in den entsprechenden Laboren erkundigt. Bislang haben wir für unsere anti-HCV NS5B-Antikörper ab35586 und ab65410 in Erfahrung bringen können, dass sie leider NICHT mit Genotyp 2a kreuzreagieren. (Derzeit haben wir anti-NS3-Antikörper verfügbar, die gut mit Genotyp 2a-Isolaten funktionieren, falls das eine Alternative für Sie sein könnte: ab13830 oder ab65407).

      https://www.abcam.com/Hepatitis-C-Virus-NS3-antibody-H23-ab13830.html (or use the following: https://www.abcam.com/Hepatitis-C-Virus-NS3-antibody-H23-ab13830.html).
      https://www.abcam.com/Hepatitis-C-Virus-NS3-antibody-8-G-2-ab65407.html (or use the following: https://www.abcam.com/Hepatitis-C-Virus-NS3-antibody-8-G-2-ab65407.html).

      Im Moment warten wir noch auf die Rückmeldung bezüglich des NS5B-Antikörpers ab100895. Wir haben in der Vergangenheit schon versucht, in Erfahrung zu bringen, mit welchen Isolaten dieser Antikörper getestet wurde (siehe Scientific Support auf dem Datenblatt). Leider konnte das Labor bisher noch keine schlüssigen Informationen liefern, aber wir bleiben dran!

      Natürlich möchte ich Ihnen kein Produkt aufdrängen, aber wenn Sie nicht länger warten wollen oder können,wäre vielleicht unser 100% Abreview Rabatt für Sie interessant: Wenn Sie ab100895 mit Genotyp 2a-Isolaten testen möchten, können Sie zum Dank einen kostenlosen Primärantikörper Ihrer Wahl von uns erhalten. Im Austausch würden wir uns über ein Abreview für ab100895 freuen. Die Bedingungen zu unserem Testprogramm können Sie unter dem folgenden Link nachlesen: https://www.abcam.com/collaborationdiscount

      Vielen Dank für Ihre Geduld.

      Ich wünsche Ihnen ein schönes Wochenende!

      Read More

      Question

      Dear technical team,   Our customer would like to purchase HCV NS5B antibody which detects 1b or 2a type.   She assume that ab100895 is right for her test but, to make sure check this please.   Thank you. Best regards,

      Read More
      Answer

      Thank you for contacting us. I am sorry for the delay in this response. ab100895 is a Mouse monoclonal antibody to Hepatitis C virus NS5B which has been used previously for ELISA and Western blotting. Unfortunately I have been unable to confirm if this antibody would recognise both 1b or 2a type. We do however have a further two antibodies against the same target: ab35586 and ab65410 which are both rabbit polyclonals which may also be suitable. These two antibodies have been tested for activity against  directly for their activity against genotypes 1a and 1b. Whilst other genotypes have not been tested being a polyclonal, it is expected to recognise other genotypes as well. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

      Read More

      Question

      Dear technical service, I have got an additional question concerning the Hepatitis C virus NS5B antibody (ab35586). Can you please tell me the genotype your antibody is based on? In the datasheet you refer to two different genotypes (entrez gene: type 1a swiss prot: 1b).

      Read More
      Answer

      Thank you for your enquiry. I can confirm that this antibody works well with both 1a and 1b genotypes and also should work with many other genotypes as it is a polyclonal raised to an almost full-length protein. I hope this information is helpful to you. Should you have any further questions, please do not hesitate to contact us.

      Read More

      Question

      ab35586 (Hepatitis C virus NS5B antibody) Will this product recognize the protein in Strain 1A?

      Read More
      Answer

      Thank you for your patience. Regarding ab35586 (Hepatitis C virus NS5B antibody), this antibody will recognize genotype 1a. Please don't hesitate to contact us again if you have further questions.

      Read More

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