Anti-V5 tag antibody [SV5-Pk1] (ab27671)

Reviews (8)Q&A (4)References (59)

Overview

  • Product name
    Anti-V5 tag antibody [SV5-Pk1]
    See all V5 tag primary antibodies
  • Description
    Mouse monoclonal [SV5-Pk1] to V5 tag
  • Host species
    Mouse
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, IHC-Fr, ELISA, IP, WB, RIAmore details
  • Immunogen

    Mice were infected with the paramyxovirus SV5, Simian-Virus 5.

  • Epitope
    ab27671 recognises a small epitope, termed Pk, present on the P/V proteins of the paramyxovirus, SV5.
  • Positive control
    • ICC/IF: CHO cells overexpressing a V5-tagged protein. WB: Mouse cell lysate - whole cell (Cortex cell line).
  • General notes

    ab27671 has been used to detect recombinant proteins, some of which include transmembrane and secreted proteins, which have been tagged with the V5 epitope.

    Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor® 488 (ab150113).

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: 7.40
    Preservative: 0.09% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Purification notes
    This antibody was purified from tissue culture supernatant by affinity chromatography.
  • Primary antibody notes
    Ab27671 has been used to detect recombinant proteins, some of which include transmembrane and secreted proteins, which have been tagged with the V5 epitope.
  • Clonality
    Monoclonal
  • Clone number
    SV5-Pk1
  • Myeloma
    Sp2/0-Ag14
  • Isotype
    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab27671 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 at an assay dependent concentration. PubMed: 21887337
Flow Cyt Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ELISA 1/1000 - 1/5000.
IP Use at an assay dependent concentration.
WB 1/1000 - 1/5000.
RIA Use at an assay dependent concentration.

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 [SV5-Pk1] (ab27671)
    Immunocytochemistry/ Immunofluorescence - Anti-V5 tag antibody [SV5-Pk1] (ab27671)Benzoubir et al PLoS One. 2015 Jun 25;10(6):e0130559. doi: 10.1371/journal.pone.0130559. eCollection 2015. Fig 1.

    γSMA is expressed in TGF-β-induced EMT.
    HuH7 (Human cell line) cells were transfected with a V5-tagged vector coding for γSMA. Cells were treated with TGF-β for 48h, and the expression of V5, γSMA and αSMA was determined by immunofluorescence analysis. Expression levels of V5 and γSMA or V5 and αSMA were determined by measuring fluorescence intensity on a linear section of the cell.

    One representative experiment is shown.

  • Western blot - Anti-V5 tag antibody [SV5-Pk1] (ab27671)
    Western blot - Anti-V5 tag antibody [SV5-Pk1] (ab27671)Flores et al PLoS One. 2016 Oct 24;11(10):e0165084. doi: 10.1371/journal.pone.0165084. eCollection 2016. Fig S2.

    Characterization of new C9orf72 dipeptide repeat antibodies.
    Rabbit polyclonal antibodies were generated against the sense strand-derived DPRs GA and GR.

    (A) Western blot of lysates from COS-7 (African green monkey kidney fibroblast-like cell line) cells transfected with the indicated vectors. Blots were serially probed with the indicated DPR antibody, then ab27671 and then actin as a loading control.

  • Immunocytochemistry/ Immunofluorescence - Anti-V5 tag antibody [SV5-Pk1] (ab27671)
    Immunocytochemistry/ Immunofluorescence - Anti-V5 tag antibody [SV5-Pk1] (ab27671)Image courtesy of an anonymous Abreview.

    ab27671 at a 1/1000 dilution staining V5 tag in CHO (Chinese hamster ovary cell line) cells overexpressing a V5-tagged protein by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton X-100. Samples were blocked with 3% BSA. The secondary used was an Alexa Fluor® 488 conjugated goat anti-mouse IgG (H+L) at a 1/1000 dilution.

  • Western blot - Anti-V5 tag antibody [SV5-Pk1] (ab27671)
    Western blot - Anti-V5 tag antibody [SV5-Pk1] (ab27671)This image is courtesy of an abreview submitted by Melina Reisenberg, CARD, KCL, London
    Anti-V5 tag antibody [SV5-Pk1] (ab27671) at 1/1000 dilution + Mouse cortex cell line lysate at 30 µg

    Secondary
    HRP-conjugated secondary antibody at 1/3000 dilution

    See Abreview

References

This product has been referenced in:
  • Douse CH  et al. Neuropathic MORC2 mutations perturb GHKL ATPase dimerization dynamics and epigenetic silencing by multiple structural mechanisms. Nat Commun 9:651 (2018). Read more (PubMed: 29440755) »
  • Helfenberger KE  et al. Subcellular distribution of ERK phosphorylation in tyrosine and threonine depends on redox status in murine lung cells. PLoS One 13:e0193022 (2018). Read more (PubMed: 29489891) »
See all 59 Publications for this product

Publishing research using ab27671? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 12 Abreviews or Q&A

Western blot abreview for Anti-V5 tag antibody [SV5-Pk1]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (NIH3T3 cells)
Gel Running Conditions
Reduced Denaturing (4%-12%)
Loading amount
20 µg
Treatment
Transfected with or without V5-tagged plasmids
Specification
NIH3T3 cells
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 Jul 25 2016

Western blot abreview for Anti-V5 tag antibody [SV5-Pk1]

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (myoblast)
Gel Running Conditions
Non-reduced Denaturing (10%)
Loading amount
40 µg
Treatment
infected with retrovirus
Specification
myoblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
Read More

Miss. Can Zhou

Verified customer

Submitted Feb 26 2016

Western blot abreview for Anti-V5 tag antibody [SV5-Pk1]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (vascular smooth muscle cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
vascular smooth muscle cell
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More

Abcam user community

Verified customer

Submitted Jan 06 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-V5 tag antibody [SV5-Pk1]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (HEK293)
Specification
HEK293
Permeabilization
Yes - 0.2% triton
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Sep 19 2013

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!

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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.

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Question

Thank you very much for your prompt response. Yes, we would like to have one vial of ab27671 (anti-V5) for replacement. Although the Mdr1 antibody did not work well, but I am very satisfied with your help. Have a great day!

Read More
Answer

Thank you for your kinds words. I am happy to help!

As requested, I have issued a free of charge replacement for one vial of ab27671.

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

We have talked on the phone and e-mailed back and forth about the Mdr1 antibody (please see the e-mails below). We tested the replacement Mdr1 (ab3366) antibody and it still did not work. To make sure that our samples have no problem, we kept trying different Mdr1 antibody from other company. As shown on the attached picture, we did see that Mdr1 level was enhanced for A549 cells after CDDP treatment but not for pc14 cells (the right picture) when detected with one Mdr1 antibody from Santa Cruz. However, when probed with ab3366 for the same membrane, we did not see the specific signal for Mdr1 (the left picture). We would like to request replacement for anti-V5 antibody (ab27671). Hopefully this antibody will work well and I don’t need to keep bothering you. Thank you very much for your help!

Read More
Answer

Thanks for your reply. I am sorry to hear the replacement vial of ab3366 did not improve things.

Just to make sure, you would like me to send along one vial of ab27671 as a replacement for this most recent faulty vial of ab3366? If this is your request I can certainly take care of that for you.

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Question

Hello, I have a technical question on the specificity of two of your antibody products, a monoclonal and a polyclonal, raised against the V5 peptide tag.   These are: 1) Anti-V5 tag antibody [SV5-Pk1] (ab27671) https://www.abcam.com/V5-tag-antibody-SV5-Pk1-ab27671.html   2) Anti-V5 tag antibody - ChIP Grade (ab9116) https://www.abcam.com/V5-tag-antibody-ChIP-Grade-ab9116.html     We are currently using these antibodies for detection of a recombinant protein which has the V5 tag at the C-terminus.   Did you or someone else test this antibody for crossreactivity with human proteins in different tissues/cell lines? If yes, what were the results?   Thank you very much!  

Read More
Answer

For ab27671 we only have data on it being used in HEK cells and for ab9116, it was used in HeLa and the western blot data is available on the public website, in the Abreview section. Both antibodies appeared to give a strong signal in cells transfected with a V5 tagged protein, but did not give a signal in the non-transfected cell lane. If there is anything else I can help you with, please let me know.

Read More

Western blot abreview for Anti-V5 tag antibody [SV5-Pk1]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Cortex cell line)
Loading amount
30 µg
Specification
Cortex cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Read More

Abcam user community

Verified customer

Submitted Nov 02 2010

Immunocytochemistry abreview for Anti-V5 tag antibody [SV5-Pk1]

 Excellent
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunocytochemistry
Sample
Chinese Hamster Cultured Cells (CHO cells overexpressing a V5-tagged protein)
Specification
CHO cells overexpressing a V5-tagged protein
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Jul 29 2009

1-10 of 12 Abreviews or Q&A

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