V5 tag properties and uses

 

The V5 epitope tag is derived from a peptide present at the C terminus of the P and V proteins of simian virus 5. It contains 14 amino acids, from N to C: GKPIPNPLLGLDST.

 

Oxford Genetics provides a range of plasmid vectors containing V5 tags in a range of orientations, to meet all your cloning requirements. Please browse through our products using the ‘Browse Plasmids’ menu. For example we provide V5 at both N and C sides of the MCS, allowing versatile positioning of your gene, we sell it with and without enzyme cleavage sites (for simple removal of the tag, if required, after protein production), and we sell it in a range of configurations with affinity tags (such as hexahistidine) to give you maximum flexibility.

 

The V5 tag is an epitope tag, meaning there are good antibodies that recognise it specifically. This makes it especially useful for immunoprecipitation, immunofluorescence and western blotting.

 

Immunoprecipitation of proteins using V5 tags: the principle

 

A typical V5 immunoprecipitation is as follows:

  1. Harvest and lyse host cells with expressed V5-tagged protein, and clarify by centrifugation





  2.  
  3.  
  4. Incubate the lysate with a slurry of gel beads bearing V5-capture antibodies, centrifuge and wash carefully.





  5.  
  6.  
  7. Elute the bound V5-tagged protein and its binding partner(s) with a buffer containing the free V5-elution peptide GKPIPNPLLGLDST.

 

Our product range

Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal V5 tags. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required.

 

The plasmid structure shown contains the V5 tag positioned downstream of the MCS, for inclusion at the C terminal of your inserted gene. The plasmid also contains puromycin resistance, driven by the ubiquitin promoter, for selection in mammalian cells, and kanamycin resistance for selection in bacteria.

 

Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.