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
    Human, mouse, or other cell types having the GKPIPNPLLGLDST tag.
  • 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

Applications

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 1/500 - 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?)

References

This product has been referenced in:
  • Amabile A  et al. Inheritable Silencing of Endogenous Genes by Hit-and-Run Targeted Epigenetic Editing. Cell 167:219-232.e14 (2016). WB . Read more (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). Read more (PubMed: 25240618) »

See all 4 Publications for this product

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

Customer reviews and Q&As

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7 reviews or Q&A

10
Votes: Highest First

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

 Good
Abreviews
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
abreview image
Username

Mr. Javier Rodriguez

Verified customer

Submitted Nov 22 2013

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 ...

Read More


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.
...

Read More

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

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
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
abreview image
Username

Mr. Javier Rodriguez

Verified customer

Submitted May 15 2013

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.

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.

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

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
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
abreview image
Username

Abcam user community

Verified customer

Submitted Mar 31 2010

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

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Oct 19 2009

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

 Average
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
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
abreview image
Username

Abcam user community

Verified customer

Submitted Dec 03 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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