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

  • 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

  • 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

  • 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

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

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

Mr. Javier Rodriguez

Verified customer

Submitted Nov 22 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 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

Mr. Javier Rodriguez

Verified customer

Submitted May 15 2013

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 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

Abcam user community

Verified customer

Submitted Mar 31 2010

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

Abcam user community

Verified customer

Submitted Oct 19 2009

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

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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