Overview

  • Product name

    Anti-Hepatitis C Virus NS3 antibody [H23]
    See all Hepatitis C Virus NS3 primary antibodies
  • Description

    Mouse monoclonal [H23] to Hepatitis C Virus NS3
  • Host species

    Mouse
  • Specificity

    Hepatitis C Virus antigen NS3 helicase domain. Reacts very well with JFH-1 strain (genotype 2a)
  • Tested applications

    Suitable for: ELISA, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Hepatitis C virus
  • Immunogen

    Recombinant full length protein (E. coli).

  • Epitope

    Helicase domain

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    H23
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab13830 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.
WB 1/1000.
IHC-P 1/50.

Tested using paraffin-embedded liver sections from HCV patients.

Target

  • Relevance

    HCV is a positive, single-stranded RNA virus in the Flaviviridae family. The genome is approximately 10,000 nucleotides and encodes a single polyprotein of about 3,000 amino acids. The polyprotein is processed by host cell and viral proteases into three major structural proteins including NS3, and several non-structural proteins necessary for viral replication. The NS3 part of the polyprotein displays three enzymatic activities: serine protease, NTPase and RNA helicase. The NS3 serine proteinase (NS3P) is a non-structural hepatitis C protein responsible for proteolytic processing of other non-structural proteins; because of this, it is also the most extensively studied protein of the Hepatitis C genome. It is responsible for proteolytic processing of the entire downstream region of the HC polyprotein, catalyzing cleavage at the NS3/NS4a, NS4a/NS4b, NS4b/NS5a, and NS5a/NS5b sites to release the mature NS3, NS4a, NS4b, NS5a, and NS5b proteins.For proper function, NS3 requires NS4a as a cofactor, but, interestingly enough, NS3 also cleaves the NS4a protein. The molecular weight of the monomer NS3P is 70 kDa.
  • Cellular localization

    Endoplasmic reticulum membrane
  • Database links

    • Alternative names

      • Hepatitis C Virus NS3 antibody
      • NS3 antibody
      • NS3P antibody
      • p70 antibody
      • Serine protease/NTPase/helicase antibody
      see all

    Images

    • ab13830 used in IHC at 1/50 dilution. Cell line used is Huh7 human hepatoma.
    • The negative control for above picture with Huh7 cells.
    • Representative Western Blot detecting Hepatitis C Virus NS3 with ab13830.

      SL209-biotin was immobilized on streptavidin agarose beads and incubated with lysates of Huh-7.5 cells infected with HCV J6/JFH-1. Retained proteins were examined by western blot.

    References

    This product has been referenced in:

    • Xu H  et al. MicroRNA-122 supports robust innate immunity in hepatocytes by targeting the RTKs/STAT3 signaling pathway. Elife 8:N/A (2019). Read more (PubMed: 30735121) »
    • Wu ZY  et al. Farnesoid X receptor agonist GW4064 indirectly inhibits HCV entry into cells via down-regulating scavenger receptor class B type I. Eur J Pharmacol 853:111-120 (2019). Read more (PubMed: 30902657) »
    See all 26 Publications for this product

    Customer reviews and Q&As

    1-4 of 4 Abreviews or Q&A

    Answer

    Thank you for your message.

    Reviewing the recommended storage details for these antibodies, I can suggest they should not be significantly affected by short term storage for a week or two at +4oC. However, I would recommend to aliquot and freeze them as soon as possible.

    We would recommend that products are stored as recommended on the datasheet as soon as they are delivered. This will ensure the product remains stable and that we can continue to provide our guarantee.

    I hope this will be helpful. If you have any further questions, please do not hesitate to contact us.

    Read More

    Answer

    Thank you for your enquiry. I am sorry it has taken some time to request the immunogen seuqences from our sources and review these for you, thank you for your patience. I can confirm the following mouse Hepatitis C Virus NS3 antibodies have been tested in wesern blot and should detect the helicase domain. I can recommend to review the online datasheets for further information: ab13830 Hepatitis C Virus NS3 antibody [H23] Mouse monoclonal Applications: ELISA, IHC-P, WB The immunogen for this mab was of the genotype 1b, the exact sequence is unknown. The epitope has been mapped to the helicase domain (between aa positions 1350 and1500). Click here (or use the following: https://www.abcam.com/index.html?datasheet=13830). ab65407 Hepatitis C Virus NS3 antibody [8 G-2] Mouse monoclonal Applications: ELISA, ICC/IF, IHC-P, WB The epitope has been mapped between amino acid positions 1340 and 1480 The antibody includes and recognises the helicase domain. Click here (or use the following: https://www.abcam.com/index.html?datasheet=65407). I am still waiting for confirmation of the immunogen from several more antibodies. I will contact you again if I have any further information that any will detect the helicase domain. I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

    Read More

    Answer

    Thank you for sending me the image of your blot. So, you have no bands (just some background) in the NS3 recombinant protein lane, and the same pattern of bands in the other lanes, including the negative control. Just to make sure, do you know that your secondary antibody is working properly? Also, have you tried in the past another Hepatitis C Virus antigen antibody with these samples?

    Read More

    Question

    BATCH NUMBER 64192 ORDER NUMBER 66184 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE Huh 20.5 (negative control) Huh BB7, GS4.1 (positive for HCV NS3) NS3 recombinant protein (positive control) Core recombinant protein (negative control) PRIMARY ANTIBODY abcam 13830 mouse monoclonal at 1:1000 and 1:500 for two hours, wash five to ten minutes. SECONDARY ANTIBODY secondary anti-mouse 1:10,000 for 2 hours and wash twice for 30 mins DETECTION METHOD ECL for 5 minutes ANTIBODY STORAGE CONDITIONS 4*C SAMPLE PREPARATION lysis buffer 300mM NaCl, 50mM Tris, 5mM EDTA, 5% Igepal, 1mMPMSF, 5ug/ml leupeptin, 30ug/ml aproptonin. lyse on ice for 10 minutes, spin and collect supernatants. AMOUNT OF PROTEIN LOADED add 10-20 ug protein with 2X SDS reducing buffer ELECTROPHORESIS/GEL CONDITIONS load and run on 10% SDSPAGE run for one and a half hours and transfer TRANSFER AND BLOCKING CONDITIONS transfer 2 hours and block using milk, tween 20 and pbs 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 WHAT STEPS HAVE YOU ALTERED? amount of primary antibodyand length of time. Hi I actually purchased this antibody the hepatitis C virus antigen and tested the Western Blot specificity using a control containing HCV NS3 positive and a negative control and did not receive a specific band in the correct size. I performed the WB at both the suggested dilution of antibody 1:1000 and at 1:500. neither blot worked. There was proper transfer of protein but too many non-specific bands and non at the specific size. Is there another antibody that I can get as a replacement or a refund?? please let me know asap, I can send you the blot as an attachment if you want to see.

    Read More
    Answer

    Thank you for your message and I'm sorry to hear that you are experiencing difficulty with ab13830. It would be helpful if you could send me an image of the blot. You mention no signal but then mention non-specific bands. How strong are these bands? What are you seeing in your negative control lanes? At this point I would like to make a few suggestions that will hopefully help you out. Try incubating with the primary at a dilution of 1:1000 overnight at 4C. Also, you mention that you are seeing non-specific bands; make sure to run a secondary only control (no primary antibody) to ensure that these bands are not due to your secondary antibody. Thanks, and I look forward to hearing from you.

    Read More

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