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    v5-tag-antibody-e10-ab53418.pdf

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Tags & Cell Markers Epitope Tags V5 Tag
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Anti-V5 tag antibody [E10] (ab53418)

  • Datasheet
Reviews (6)Q&A (2)References (5)

Product price, shipping and contact information

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Immunocytochemistry/ Immunofluorescence - Anti-V5 tag antibody [E10] (ab53418)
  • Western blot - Anti-V5 tag antibody [E10] (ab53418)

Key features and details

  • Mouse monoclonal [E10] to V5 tag
  • Suitable for: ICC/IF, IHC-P, WB, ELISA, IHC-Fr
  • Reacts with: Species independent
  • Isotype: IgG1

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Overview

  • Product name

    Anti-V5 tag antibody [E10]
    See all V5 tag primary antibodies
  • Description

    Mouse monoclonal [E10] to V5 tag
  • Host species

    Mouse
  • Specificity

    ab53418 detects (GKPIPNPLLGLDST) V5 tag fusion protein.
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, ELISA, IHC-Frmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Synthetic peptide:

    GKPIPNPLLGLDST

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

    • 293 cells untransfected and transfected with lacZ-V5-tagged.

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 long term.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituent: 0.0268% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    ab53418 was affinity purified from culture supernatant of hybridoma cells grown in a bioreactor.
  • Clonality

    Monoclonal
  • Clone number

    E10
  • Isotype

    IgG1
  • Research areas

    • Tags & Cell Markers
    • Epitope Tags
    • V5 Tag

Associated products

  • Compatible Secondaries

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

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab53418 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
ICC/IF (1)
1/500 - 1/2000.
IHC-P
Use at an assay dependent concentration.
WB (5)
1/1000 - 1/3000. Detects a band of approximately 130 kDa.
ELISA
1/5000 - 1/10000.
IHC-Fr
1/500 - 1/2000.
Notes
ICC/IF
1/500 - 1/2000.
IHC-P
Use at an assay dependent concentration.
WB
1/1000 - 1/3000. Detects a band of approximately 130 kDa.
ELISA
1/5000 - 1/10000.
IHC-Fr
1/500 - 1/2000.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-V5 tag antibody [E10] (ab53418)
    Immunocytochemistry/ Immunofluorescence - Anti-V5 tag antibody [E10] (ab53418)This image is courtesy of an Abreview submitted by Javier Rodriguez

    ab53418 staining V5 tag in Mouse B16F1 melanoma cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Tween 20 and blocked with 3% BSA for 1 hour at 23°C. Samples were incubated with primary antibody (1/500 in 1% BSA + 0.5% Tween 20 + 1% Donkey serum) for 1 hour at 21°C. An Alexa Fluor® 555-conjugated Donkey anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

    Melonma cells expressing GFP are also expressing GFP (AF55) due to transfected cell line with GFP-V5.

    See Abreview

  • Western blot - Anti-V5 tag antibody [E10] (ab53418)
    Western blot - Anti-V5 tag antibody [E10] (ab53418)
    All lanes : Anti-V5 tag antibody [E10] (ab53418) at 1/1000 dilution

    Lane 1 : 293 cells
    untransfected (control)
    Lane 2 : 293 cells
    transfected
    with lacZ-V5-tagged.

    Lysates/proteins at 30 µg per lane.

    Observed band size: 130 kDa
    why is the actual band size different from the predicted?

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (5)

    Publishing research using ab53418? Please let us know so that we can cite the reference in this datasheet.

    ab53418 has been referenced in 5 publications.

    • Mora Gallardo C  et al. Dido3-dependent SFPQ recruitment maintains efficiency in mammalian alternative splicing. Nucleic Acids Res 47:5381-5394 (2019). PubMed: 30931476
    • Amabile A  et al. Inheritable Silencing of Endogenous Genes by Hit-and-Run Targeted Epigenetic Editing. Cell 167:219-232.e14 (2016). WB . PubMed: 27662090
    • Wang J  et al. Sumoylation modulates 20-hydroxyecdysone signaling by maintaining USP protein levels in Drosophila. Insect Biochem Mol Biol 54C:80-88 (2014). PubMed: 25240618
    • Cho SW  et al. Novel role of hnRNP-A2/B1 in modulating aryl hydrocarbon receptor ligand sensitivity. Arch Toxicol N/A:N/A (2014). PubMed: 25224401
    • Zhang F  et al. The African swine fever virus DP71L protein recruits the protein phosphatase 1 catalytic subunit to dephosphorylate eIF2alpha and inhibits CHOP induction but is dispensable for these activities during virus infection. J Virol 84:10681-9 (2010). PubMed: 20702639

    Customer reviews and Q&As

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

    1-8 of 8 Abreviews or Q&A

    Western blot abreview for Anti-V5 tag antibody [E10]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK-293T)
    Gel Running Conditions
    Reduced Denaturing (8-13%)
    Loading amount
    35 µg
    Specification
    HEK-293T
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Dr. Jeeho Kim

    Verified customer

    Submitted Jan 09 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-V5 tag antibody [E10]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
    Sample
    Mouse Cell (B16F1 melanoma cell line)
    Specification
    B16F1 melanoma cell line
    Permeabilization
    Yes - 0,5% tween 20
    Fixative
    Paraformaldehyde
    Read More

    Mr. Javier Rodriguez

    Verified customer

    Submitted Nov 22 2013

    Question

    I would like to use the antibody for IHC and IF. I can see the starting dilution you recommend using when applying the Ab for those techniques, but I did not see any clue or control for those applications. Do you think this antibody is recommended for fluorescense imaging, or is better to try another one as the ab9113 or ab1209? Please if you have further information about the best protocol that fits the Ab conditions as Acidic buffer, retrieval conditions etc, It would be a great help for me.

    Read More

    Abcam community

    Verified customer

    Asked on May 21 2013

    Answer


    Since this antibody is against a V5 tag which isn't found naturally in tissues, the only positive control we can recommend are cells/tissues transfected with a V5 tagged protein. A negative control can simply be untransfected cells/tissues.

    Choosing to do a fluorescent detection rather than an enzymatic detection depends solely on the secondary antibody used and on the application you are using it in. The choice of primary antibody will not affect the fluorescence imaging therefore neither one of ab53418, ab9113 and ab1209 are recommended above the other for IF. In general IHC on paraffin sections uses a enzymatic detection whereas IHC on frozen sections uses a fluorescent detection although the detection methods are interchangeable.

    I would recommend using a citrate buffer pH6 for the antigen retrieval and blocking with 10% serum or 5% BSA for 1 h at RT followed by an overnight incubation at 4ºC with 1/1000 primary antibody.

    Read More

    Abcam Scientific Support

    Answered on May 21 2013

    Western blot abreview for Anti-V5 tag antibody [E10]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Whole cell lysate and conditioned media from the B)
    Loading amount
    1e+006 cells
    Specification
    Whole cell lysate and conditioned media from the B
    Gel Running Conditions
    Reduced Denaturing (10% Acrylamide)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Mr. Javier Rodriguez

    Verified customer

    Submitted May 15 2013

    Question

    I would like to order an anti-V5 tag which is either mouse or rabbit monoclonal for use in western blots. I think my best option is ab53418. Am I right? Or could you suggest a better/alternative option.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 22 2012

    Answer

    Thank you for contacting Abcam.

    I think ab53418 would be an excellent choice for you and under our Abpromise We guarantee that this antibody will work in western blot.

    If there is anything else I can help you with, please let me know.

    Read More

    Abcam Scientific Support

    Answered on Feb 22 2012

    Western blot abreview for Anti-V5 tag antibody [E10]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HepG2)
    Loading amount
    30 µg
    Specification
    HepG2
    Gel Running Conditions
    Reduced Denaturing (4-12% Nupage)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 31 2010

    Western blot abreview for Anti-V5 tag antibody [E10]

    Good
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293T)
    Specification
    293T
    Gel Running Conditions
    Reduced Denaturing (10% Bis-Tris Gel)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 19 2009

    Western blot abreview for Anti-V5 tag antibody [E10]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (PC3 Cells (Prostate cancer line))
    Loading amount
    50 µg
    Specification
    PC3 Cells (Prostate cancer line)
    Gel Running Conditions
    Reduced Denaturing
    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 Dec 03 2007

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