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

Reviews (8)Q&A (4)References (72)

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

  • Species reactivity

    Reacts with: Species independent

  • 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

    • WB: Transfected NIH/3T3 and COS-7 cell lysates. ICC/IF: CHO cells overexpressing a V5-tagged protein.

  • 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.

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)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)
    All lanes : Anti-V5 tag antibody [SV5-Pk1] (ab27671) at 1/1000 dilution

    Lanes 1-2 : Transfected NIH/3T3 cell lysate
    Lane 3 : Not transfected NIH/3T3 cell lysate

    Secondary
    All lanes : HRP-conjugated rabbit polyclonal Anti-mouse

    Developed using the ECL technique.

    Exposure time: 1 second


    Cells were transfected with a V5-tagged HSPB8 (MW:~25kD) plasmid, which expressed a V5-tagged protein with molecular weight of approximately 28kD.

    See Abreview

  • 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. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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.

References

This product has been referenced in:

  • Salazar VS  et al. Reactivation of a developmental Bmp2 signaling center is required for therapeutic control of the murine periosteal niche. Elife 8:N/A (2019). Read more (PubMed: 30735122) »
  • Coelho JPL  et al. A network of chaperones prevents and detects failures in membrane protein lipid bilayer integration. Nat Commun 10:672 (2019). Read more (PubMed: 30737405) »
See all 72 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

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

1-10 of 12 Abreviews or Q&A

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

 Excellent
Abreviews
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
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
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
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!

Read More
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.

Read More

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.

Read More

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
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
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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com