Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online

Anti-V5 tag antibody (ab9116)

Reviews (13)Q&A (18)References (129)
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

    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

    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

    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

    11-20 of 31 Abreviews or Q&A

    Immunocytochemistry abreview for Anti-V5 tag antibody - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (Panc1 cancer cell)
    Specification
    Panc1 cancer cell
    Fixative
    Acetone
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 20°C
    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 Mar 23 2012

    Question

    Hi, I would like to know if I could benefit from a similar offer for antibodies: ab9116 (ChIP grade anti-V5) for Intracellular Flow Cytometry. ab131536 (rabbit anti-EphrinB2) for Flow Cytometry/Intracellular Flow Cytometry.

    Read More
    Answer

    I am very pleased to hear you would like to accept our offer and test these products in flow cytometry. The code will give you the VALUE of each product off an order before the expiration date. To redeem this offer, please submit an Abreview for flow cytometry. The code is inactive until included in your submitted Abreview so be sure to add it to the Additional Comments section. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/AbTrial

    Read More

    Question

    I spoke to you last Friday regarding the Rabbit polyclonal antibody V5(ab9116). The antibody failed in Western blot, IF and CO-IPS. You said there will be a free replacement for us. The replacement should be ab27671 mouse monoclonal antibody.  I forwarded you the purchase confirmation below. Please let me know if you need more information. Thanks!

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. 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. 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.

    Read More

    Question

    Thank you for replying to me.
    I cloned the V5 tag at the N-terminal extremity of my protein and in
    frame. I placed only one tag.
    I added an HA tag instead and I was able to detect my protein.
    Best regards,

    Read More
    Answer

    Thank you for your updates. I am very pleased to hear that you obtained good results.

    Please do let me know how you wish to proceed with this enquiry.

    If you need any further assistance in the future, please do not hesitate to contact me.

    Read More

    Question

    Hello! I am studying different domains of a given protein by generating truncated mutants. I tagged all the mutants with the V5 tag. For my western blots, I block the membrane 30min in TBST+milk, I incubate overnight at 4°C with anti-V5 diluted 1/5,000 and 1h at RT with anti-rabbit diluted 1/5,000. I wash 4x5min in TBST. It works very well with most of my mutants but not with the three smaller ones between 10 and 25 kDa for which I do not get any signal. I tried with a higher number of cells, used a 20microns nitrocellulose membrane and decreased transfer time, still no detection. I also tried to add MG132 to my cell culture after transfection in case of quick turn-over but still no result. I performed immunofluorescence but no signal with my small proteins and strong signal with heavier ones. Is the tag structure unstable with small proteins? Thank you in advance for all the help you might bring to me.

    Read More
    Answer

    Thank you for your enquiry regarding ab9116.

    We are very sorry about the delay in response but have been trying to get in touch with the originator of this product.

    At this stage, it is difficult to identify the source of the problem. We would suggest loading more protein in the lanes and/or increasing the working concentration of this antibody to 1 ug/ml.

    - Could you please confirm if you have have run a gel to check the proteins are being expressed?

    - Where is the location of the tag within the reading frame of the target protein?

    - How many copies of tags have been inserted?

    I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

    Read More

    Question

    Dear technical support:

    This customer raised an inquiry about ab9116 (Anti-V5 tag antibody - ChIP Grade), and this customer used this antibody conduct the western blotassay with human sample, and the data shows multiple bands and no major bean, because this antibody is already out of our warranty period, therefore he only wants to ask for your kindly help to offer any suggestion to him. I attach the western blot data and the WB questionnaire in this letter, could you please help this customer to solve this problem.

    Thanks for your kindly assistance.

    Best regards,

    Read More
    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    I would like to reassure you that ab9116 is tested and covered by our 6 month guarantee for use in WB. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

    Reviewing this case, I would like to offer some suggestions to help optimize the results. I would also appreciate if you can confirm some further details:

    1. According to our records for the order number provided, this was purchased in March. Our guarantee is 6 months, so this would still be covered by the Abpromise guarantee. Could you confirm this was the correct order number?

    2.There are inherent difficulties with antibody detection of recombinant tagged proteins that I can recommend considering. This may be a biological artifact rather than a problem with the antibody. Could you confirm that the protein transfected is full length and includes the immunogen sequence? As the recombinant is tagged, it is possible that the tag is preventing access to the antibody. Could you confirm at which region of the protein the tag has been placed?

    3. Could you confirm how the stability of transfection has been assessed? The sample may contain different fragment sizes.

    4. Has the quality of the sample been assessedwith loading control?

    5. I can suggest to run the samples on a reducing gel. This will ensure the sample is in the correct conformation to run at the correct molecular weight.

    6. I can recommend to try a lower concentration of antibody to reduce the sginal. Try 1:10 000.

    7. Is the current vial of secondary antibody working well with other primary antibodies? It may be that the concentration needs to be reduced in order to optimize the results. What are the results of the no primary control?

    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

    Question

    Inquiry: Hi, as a point of information I would say it is misleading to quote the following reference on your site as confirmation of use of ab15828 in ChIP applications as the Ab in question is, as far as I can tell, only used as a negative control in a pre-clear prior to ChIP with other antibodies. The reason I have been looking into this is that I am trying to decide which is the better antibody between 9616 and 15828 for ChIP applications using v5 epitope. Any suggestions would be most welcome. http://www.sciencedirect.com/science/article/pii/S0012160608011986

    Read More
    Answer

    Thank you for your inquiry.
    I would like to reassure you that both antibodies are tested and guaranteed for CHIP.
    I can confirm that ab15828 was tested by our lab in house for CHIP and therefore I would recommend to use this antibody.
    If you find the publications for ab9116 more convincing, I would like to confirm again that this antibody is also guaranteed for CHIP by our Abpromise.
    ab15828 has more than one CHIP publication and therefore we believe that it is also valuable information if a publication states that this antibody can be used as a negative control for other antibodies.
    I hope this information is helpful. Please do not hesitate to contact me again with any further questions.

    Read More

    Question

    WB with V5-tagged protein, expected band at 52 kDa
    lot GR20095-3 worked (crtl: no band, samples: 52 kDa)
    new lot GR61535-2: 52 kDa band, but also band at48, 40 and about 30kDa, sees these extra bands also in control samples, thus non-specific
    intensity of bands is equally strong

    Ab: 1/10'000 4h 4 degree
    2nd: OK
    block: 5 % milk

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    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 ab15828 and ab27671. The order will ship on Monday for Tuesday delivery as you had requested.

    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.

    Read More

    Question

    I appreciate a lot your informative and helpful reply. The customer decided to proceed with the abreview promotion of ab9116 as you mentioned for IHC-Fr (following our correspondence below). Please issue the discount code ASAP, because the customer would like to add this antibody to our order that will be shipped tomorrow (this Friday). Thanks in advance for your reply. Happy Holidays,

    Read More
    Answer

    DISCOUNT CODE: XXXXXX Expiration date: 20-Apr-2012 I am very pleased to hear you would like to accept our offer and test ab9116 in IHC-FR (4% PFA fixed Drosophila sections). This code will give you 1 free [PRIMARY ANTIBODY] before the expiration date. To redeem this offer, please submit an Abreview for IHC-FR and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

    Read More

    Question

    Thank you for the mail. As I mentioned in my previous mail the customer works with Drosophila BRAIN (fresh brain) that is fixed with 4% paraformaldehyde for 10 min. The customer does not perform any antigen retrieval. As this customer explained me this type of the staining that he performs is very similar to the staining of the cells.   Please advise if the following antibodies will be covered by Abcam's abpromise or you advise different choice. If these are not covered by Abpromise will it be possible to proceed with the Abreview promotion (the most urgent is ab98795). https://www.abcam.com/V5-tag-antibody-ChIP-Grade-ab9116.html https://www.abcam.com/V5-tag-antibody-FITC-ab1274.html https://www.abcam.com/V5-tag-antibody-ab84164.html https://www.abcam.com/HA-tag-antibody-ab9134.html https://www.abcam.com/Donkey-polyclonal-Secondary-Antibody-to-Mouse-IgG-H-L-DyLight-550-pre-adsorbed-ab98795.html Thank you an advance.   Kind Regards,  

    Read More
    Answer

    Thank you for contacting us. ab1274, ab9116 and ab98795 are not covered under Abpromise guarantee as per customer request. I am including other product under IHC-Fr application with PFA as fixative. Ab98795 is a secondary antibody; Abreview discount code is not valid for this product. However if customer provide us Abreview we will assign the Abpoints by manually publishing the Abreview. You can forward the Abreview to me. The Abreview discount code for ab1274 and ab9116 can be provided however customer needs to explain how they have expressed tag conjugated protein in drosophila; please also forward the end user details. Please be also advised that ab1274, ab9116, ab9134, ab84164 are the antibodies against tags. These tags are not naturally present in any cells. These products can only be used in detecting the tag conjugated recombinant proteins that are expressed in tissues or cells. 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

    11-20 of 31 Abreviews or Q&A

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