Name: Anti-V5 tag antibody [E10]
SKU: ab53418
Rating: 4 (5 reviews)

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Primary

Secondary

This product Mouse Anti-V5 tag antibody [E10] (ab53418)
ICC/IF, IHC-P, WB, ELISA, IHC-Fr

Goat Anti-Mouse IgG H&L (HRP) (ab205719)
WB, IP, ELISA, IHC-P

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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

Reacts with: Species independent
Immunogen

Synthetic peptide:
GKPIPNPLLGLDST

Run BLAST with Run BLAST with
Positive control
- 293 cells untransfected and transfected with lacZ-V5-tagged.

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

Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
- Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)

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.

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

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)

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?

Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Datasheet

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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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

Question

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 fluorescense imaging, or is better to try another one as the ab9113 or ab1209? Please if you have further information about the best protocol that fits the Ab conditions as Acidic buffer, retrieval conditions etc, It would be a great help for me.

Abcam community

Verified customer

Asked on May 21 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.

Abcam Scientific Support

Answered on May 21 2013

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

Question

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.

Abcam community

Verified customer

Asked on Feb 22 2012

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.

Abcam Scientific Support

Answered on Feb 22 2012

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

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

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

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Anti-V5 tag antibody [E10] (ab53418)

Overview

Properties

Associated products

Applications

Target

Images

Protocols

Datasheets and documents

References

This product has been referenced in:

Customer reviews and Q&As

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

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

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

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

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