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  1. Link

    v5-tag-antibody-ab9116.pdf

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Tags & Cell Markers Epitope Tags V5 Tag
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Anti-V5 tag antibody (ab9116)

  • Datasheet
  • SDS
Reviews (13)Q&A (18)References (129)

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Western blot - Anti-V5 tag antibody (ab9116)

    Key features and details

    • Rabbit polyclonal to V5 tag
    • Suitable for: WB, ELISA, IP, IHC-P
    • Reacts with: Species independent
    • Isotype: IgG

    Get better batch-to-batch reproducibility with a recombinant antibody

    Product image
    Anti-V5 tag antibody [SV5-P-K] (ab206566)
    • Research with confidence – consistent and reproducible results with every batch
    • Long-term and scalable supply – powered by recombinant technology for fast production
    • Success from the first experiment – confirmed specificity through extensive validation
    • Ethical standards compliant – production is animal-free

    Overview

    • Product name

      Anti-V5 tag antibody
      See all V5 tag primary antibodies
    • Description

      Rabbit polyclonal to V5 tag
    • Host species

      Rabbit
    • Specificity

      ELISA: V5 tag antibody - ChIP Grade (ab9116) specificity was verified by ELISA against the peptide conjugated to BSA (GKPIPNPLLGLDST-BSA). A 1:50,000 dilution of the antibody gave an O.D.=1.0 in a 15 minute reaction using HRP- conjugated goat anti rabbit (IgG) at 1:10000 and TMB as the substrate. Appropriate specificity controls were run.

    • Tested applications

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

      Reacts with: Species independent
    • Immunogen

      Synthetic peptide corresponding to Parainfluenza virus 5 V5 tag aa 50-150 conjugated to keyhole limpet haemocyanin. (V5 epitope)
      Database link: P11208
      (Peptide available as ab15829)

      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
    • 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

    • 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: 7.20
      Preservative: 0.1% Sodium azide
      Constituent: PBS
    • Concentration information loading...
    • Purity

      Immunogen affinity purified
    • Purification notes

      Antibodies were immunoaffinity purified using the peptide immobilized on a solid phase. Antibody concentration was determined by extinction coefficient : O.D. 1.4 at 280nM equals 1.0 mg .
    • Clonality

      Polyclonal
    • Isotype

      IgG
    • Light chain type

      unknown
    • Research areas

      • Tags & Cell Markers
      • Epitope Tags
      • V5 Tag

    Associated products

    • ChIP Related Products

      • Rabbit Anti-Mouse IgG H&L (ab46540)
      • ChIP Kit (ab500)
    • Compatible Secondaries

      • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
      • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Immunizing Peptide (Blocking)

      • V5 tag peptide (ab15829)
    • Isotype control

      • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
    • Positive Controls

      • E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395)

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab9116 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
    WB (7)
    Use a concentration of 0.05 - 0.5 µg/ml. Can be blocked with V5 tag peptide (ab15829).
    ELISA
    Use at an assay dependent concentration.
    IP (2)
    Use at an assay dependent concentration.
    IHC-P
    Use a concentration of 0.5 - 5 µg/ml.
    Notes
    WB
    Use a concentration of 0.05 - 0.5 µg/ml. Can be blocked with V5 tag peptide (ab15829).
    ELISA
    Use at an assay dependent concentration.
    IP
    Use at an assay dependent concentration.
    IHC-P
    Use a concentration of 0.5 - 5 µg/ml.

    Target

    • Relevance

      The V5 epitope tag is derived from a small epitope (Pk) present on the P and V proteins of the paramyxovirus of simian virus 5 (SV5). The V5 tag is usually used with all 14 amino acids (GKPIPNPLLGLDST), although it has also been used with a shorter 9 amino acid sequence (IPNPLLGLD).
    • Alternative names

      • GKPIPNPLLGLDST epitope tag antibody
      • GKPIPNPLLGLDST tag antibody
      • Protein Rev antibody
      • Regulator of expression of viral proteins antibody
      • rev antibody
      • V5 epitope tag antibody
      see all

    Images

    • Western blot - Anti-V5 tag antibody (ab9116)
      Western blot - Anti-V5 tag antibody (ab9116)This image is courtesy of an anonymous Abreview
      All lanes : Anti-V5 tag antibody (ab9116) at 1/5000 dilution

      Lanes 1-3 : Cells transfected with v5-tagged protein.
      Lanes 4-5 : Untransfected cells.

      Secondary
      Lanes 1-3 : Anti-rabbit Ig-HRP at 1/20000 dilution
      Lanes 4-5 : Anti-rabbit Ig-HRP at 1/20000 dilution


      Mouse ES cells lysate.

      See Abreview

    Protocols

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

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (129)

    Publishing research using ab9116? Please let us know so that we can cite the reference in this datasheet.

    ab9116 has been referenced in 129 publications.

    • Harland LTG  et al. The T-box transcription factor Eomesodermin governs haemogenic competence of yolk sac mesodermal progenitors. Nat Cell Biol 23:61-74 (2021). PubMed: 33420489
    • Rashnonejad A  et al. Designed U7 snRNAs inhibit DUX4 expression and improve FSHD-associated outcomes in DUX4 overexpressing cells and FSHD patient myotubes. Mol Ther Nucleic Acids 23:476-486 (2021). PubMed: 33510937
    • Nimmanon T  et al. The ZIP6/ZIP10 heteromer is essential for the zinc-mediated trigger of mitosis. Cell Mol Life Sci 78:1781-1798 (2021). PubMed: 32797246
    • Gestuveo RJ  et al. Analysis of Zika virus capsid-Aedes aegypti mosquito interactome reveals pro-viral host factors critical for establishing infection. Nat Commun 12:2766 (2021). PubMed: 33986255
    • Hollingsworth LR  et al. DPP9 sequesters the C terminus of NLRP1 to repress inflammasome activation. Nature 592:778-783 (2021). PubMed: 33731932
    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 31 Abreviews or Q&A

    Immunoprecipitation abreview for Anti-V5 tag antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Mouse Cell lysate - whole cell (mouse neural stem cell)
    Total protein in input
    500 µg
    Immuno-precipitation step
    Protein A/G
    Specification
    mouse neural stem cell
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 15 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-V5 tag antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2OS cells)
    Permeabilization
    Yes - 0.2% Triton X-100
    Specification
    U2OS cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 06 2017

    Western blot abreview for Anti-V5 tag antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (H1299)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    20 µg
    Treatment
    transfected with V-5 p53
    Specification
    H1299
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 28 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-V5 tag antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (human u2os cells)
    Permeabilization
    Yes - 0.1% triton
    Specification
    human u2os cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 13 2016

    Western blot abreview for Anti-V5 tag antibody - ChIP Grade

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (NIH/3T3 fibroblasts with V5-tag constructs)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    NIH/3T3 fibroblasts with V5-tag constructs
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Pawel Niewiadomski

    Verified customer

    Submitted Feb 09 2016

    Western blot abreview for Anti-V5 tag antibody - ChIP Grade

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    15 µg
    Specification
    HEK293
    Blocking step
    Thermo Scientific #EN37543 Starting Block TBST formulation as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 22 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-V5 tag antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
    Sample
    Monkey Cell (cos7 cell)
    Specification
    cos7 cell
    Permeabilization
    Yes - 0.2% tritonx
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 19 2013

    Western blot abreview for Anti-V5 tag antibody - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Recombinant protein (293 cells transfected with a V5-tagged construct)
    Loading amount
    2000 cells
    Specification
    293 cells transfected with a V5-tagged construct
    Gel Running Conditions
    Reduced Denaturing (10% AA gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 19 2012

    Question

    Pouvez-vous me donner des indications sur la dilution conseillée avec l’anticorps anti-V5 (ab9116) pour faire du ChIP-Seq ?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 04 2013

    Answer


    Nous n’avons pas testé cet anticorps dans nos laboratoires et donc nous n’avons pas de dilution spécifique à recommander pour ab9116 appart les informations dans la publication suivante :
    Sugiyama T et al. SHREC, an effector complex for heterochromatic transcriptional silencing. Cell 128:491-504 (2007).
    Cependant, je vous recommande d’utiliser 3-5 mg d'anticorps pour chaque 25-35 mg de monosomes utilisés mais comme de nombreuses techniques, il sera essentiel d'optimiser la quantité d'anticorps.

    Read More

    Abcam Scientific Support

    Answered on Feb 04 2013

    Question

    This antibody is staining all nuclei in mouse brain sections, even those not expressing the tagged protein. Western blot shows a band of 55 kDa in all samples. Several different blocking steps were tried, the secondary is working well with another primary antibody. Please help with the background problem!

    Read More

    Abcam community

    Verified customer

    Asked on Nov 01 2012

    Answer

    Thank you for calling today, and I'm sorry to hear that our V5 antibody is giving you so much trouble!

    As we discussed over the phone, I'm unsure of why the antibody is detecting that band in your samples and all of the protocol modifications you've made are what I would recommend. It may be, as you suggested, that the antibody is cross-reacting with a mouse brain protein. I'll ensure that we look into this issue further, and if we believe there is a larger problem with the product, we will remove it from the catalog.

    I've arranged a free of charge vial of ab9113 (chicken polyclonal to V5) , and you should receive this tomorrow. The order number for this free of charge order is #######.

    Please let me know how the replacement antibody works for you - I want to make sure you have something that works in the end. Have a great day, and best of luck in your research!

    Read More

    Abcam Scientific Support

    Answered on Nov 01 2012

    1-10 of 31 Abreviews or Q&A

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