Overview

  • Product name
    Anti-V5 tag antibody - ChIP Grade
    See all V5 tag primary antibodies
  • Description
    Rabbit polyclonal to V5 tag - ChIP Grade
  • Host species
    Rabbit
  • Specificity
    ELISA: Antibody 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: ICC/IF, ChIP, WB, ELISA, IP, ChIP/Chip, IHC-P, ICCmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    GKPIPNPLLGLDST (V5 epitope) conjugated with KLH.

  • Positive control
    • E. coli Positive Control (Escherichia coli ) Whole Cell Lysate (ab5395) can be used as a positive control in WB. Ab5395

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

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
ICC/IF Use a concentration of 2 - 10 µg/ml. PubMed: 18575576
ChIP Use at an assay dependent concentration. PubMed: 17289569
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.
ChIP/Chip Use at an assay dependent concentration. PubMed: 17289569
IHC-P Use a concentration of 0.5 - 5 µg/ml.
ICC Use at an assay dependent concentration. PubMed: 17012257

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

  • All lanes : Anti-V5 tag antibody - ChIP Grade (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

  • ab9116 staining V5 tag in the Human U2OS cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 3% serum for 1 hour at 4°C. Samples were incubated with primary antibody (1/300) for 1 hour at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody (1/300).

    See Abreview

  • ChIP qPCR: The graph on the left represents the input chromatin of PC3-EMT14-OVOL2 relative to empty vector (EV) control. The graph on the right depicts the ChIP DNA using V5 antibody (ab9116). The V5 epitope was included at the C-terminus of the expressed OVOL2. Results were normalized to input controls and graphs are relative to EV. Graphs show mean +/- sem.

  • ab9116 at 1/1000 dilution staining V5 in V5-Pgc transfected Drosophila Kc cells by Immunocytochemistry/ Immunofluorescence. Kc cells were fixed in 4% formaldehyde for 10 minutes, washed in PBS-Tween 20 (0.05%) and blocked in 5% milk in PBS-Tween 20 for 1 hour. Cells were incubated with primary antibody at 4°C overnight and the secondary antibody at room temperature for 3 hours.

References

This product has been referenced in:
  • He Y  et al. Androgen receptor splice variants bind to constitutively open chromatin and promote abiraterone-resistant growth of prostate cancer. Nucleic Acids Res 46:1895-1911 (2018). Read more (PubMed: 29309643) »
  • Holland SJ  et al. Expansions, diversification, and interindividual copy number variations of AID/APOBEC family cytidine deaminase genes in lampreys. Proc Natl Acad Sci U S A 115:E3211-E3220 (2018). Read more (PubMed: 29555777) »
See all 75 Publications for this product

Customer reviews and Q&As

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1-10 of 13 Abreviews

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

Abcam user community

Verified customer

Submitted Jun 15 2018

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

Abcam user community

Verified customer

Submitted Sep 06 2017

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

Abcam user community

Verified customer

Submitted Aug 28 2017

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

Abcam user community

Verified customer

Submitted Jun 13 2016

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

Pawel Niewiadomski

Verified customer

Submitted Feb 09 2016

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

Abcam user community

Verified customer

Submitted Apr 22 2015

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

Abcam user community

Verified customer

Submitted Sep 19 2013

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

Abcam user community

Verified customer

Submitted Nov 19 2012

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

Abcam user community

Verified customer

Submitted Mar 23 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T cells)
Total protein in input
1000 µg
Specification
293T cells
Immuno-precipitation step
Protein A/G

Abcam user community

Verified customer

Submitted Oct 23 2009

1-10 of 13 Abreviews

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