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V5 Tag: Properties, origin, specifications, antibodies & Nanobodies

Posted by Christoph Eckert on Sep 23, 2020 2:09:32 PM

V5 tag is a short peptide tag for detection and purification of proteins. The V5 tag can be fused/cloned to a recombinant protein and detected in ELISA, flow cytometry, immunoprecipitation, immunofluorescence, and Western blotting with antibodies and Nanobodies.

  1. 3D structure of V5 tag
  2. Origin
  3. Properties
  4. Size of V5 tag
  5. Specifications of the V5 tag
  6. V5 epitope tag sequences
  7. Applications
  8. V5 tag vs. Flag tag
  9. V5 tag vs. HA tag
  10. How to elute V5 tag?
  11. V5 tag plasmids
  12. V5 tag antibodies and Nanobodies - Test sample request
  13. V5 tag beads
  14. References

 

1. 3D structure of V5 tag

V5

2. Origin

The V5 tag derived from the P and V protein of the simian virus 5 (SV5, a paramyxovirus), amino acid residues 95 to 108 of RNA polymerase alpha subunit. First referenced in Hanke et al, Journal of General Virology 19921.

 

3. Properties

The V5 tag contains 14 amino acids (aa) and the sequence is GKPIPNPLLGLDST. There is also a shorter version with 9 aa (IPNPLLGLD) which is rarely used.

 

4. Size of V5 tag

Number of amino acids: 14

Molecular weight (MW): 1421.66 Da

 

5. Specifications of the V5 tag

Theoretical isoelectric point (pI): 5.84

Total number of negatively charged residues (Asp + Glu): 1

Total number of positively charged residues (Arg + Lys): 1

 

6. V5 epitope tag sequences

V5 tag amino acid sequence:

GKPIPNPLLGLDST

 

V5 tag DNA sequence:

GGT AAG CCT ATC CCT AAC CCT CTC CTC GGT CTC GAT TCT ACG

Please note, the mentioned sequence is for mammals and insects. The sequence is not codon optimized for other organisms.

 

7. Applications

The most common applications of V5-tagged proteins are ELISA, flow cytometry, immunofluorescence (IF), immunoprecipitation (IP), protein purification, and Western blotting.

 

8. V5 tag vs. Flag tag

Here is a comparison of V5 tag and Flag tag

 

V5 tag

Flag tag

Origin

P and V protein of the simian virus 5

artificial design

3D Structure

V5 Flag-tag2

Amino acid sequence

GKPIPNPLLGLDST

DYKDDDDK

Number of amino acids

14

8

Molecular weight (in Da)

1421.66

1012.98

Theoretical isoelectric point (pI)

5.84

3.97

Total number of negatively charged residues (Asp + Glu)

1

5

Total number of positively charged residues (Arg + Lys)

1

2

 

9. V5 tag vs. HA tag

Here is a comparison of V5 tag and HA tag

 

V5 tag

HA tag

Origin

P and V protein of the simian virus 5

Human influenza hemagglutinin (HA)is derived from human influenza virus

3D Structure

V5 HA_2

Amino acid sequence

GKPIPNPLLGLDST

YPYDVPDYA

Number of amino acids

14

9

Molecular weight (in Da)

1421.66

1102.17

Theoretical isoelectric point (pI)

5.84

3.56

Total number of negatively charged residues (Asp + Glu)

1

2

Total number of positively charged residues (Arg + Lys)

1

0

 

10. How to elute V5 tag?

Elution of V5-tagged proteins is normally performed by glycine or citric acid or by competitive elution with V5 peptide (GKPIPNPLLGLDST).

 

11. V5 tag plasmids

There are commercially available plasmids for expression with V5 tag for mammalian cells, Drosophila, etc. from ThermoFisher and Merck (Sigma Aldrich).

 

12. V5 tag antibodies and Nanobodies

There are different polyclonal and monoclonal antibodies and one Nanobody against V5 commercially available. For immunoprecipitation ChromoTek’s, the anti V5 Nanobody V5-Trap is recommended as Nanobodies have lower background compared to antibodies: No dissociating heavy and light antibody chains.

You can request a free test sample of the V5-Trap here.

13. V5 tag beads

Chromotek offers V5 Nanobodies conjugated to beads for immunoprecipitation and unconjugated V5 Nanobodies/ VHHs:

V5-Trap Agarose: anti-V5 Nanobody conjugated to agarose beads

V5-Trap Magnetic Agarose: anti-V5 Nanobody conjugated to magnetic agarose beads

V5 VHH: anti-V5 Nanobody, unconjugated

 

14. Reference

  • Tomas Hanke, Paul Szawlowski and Richard E. Randall; Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using tag-specific monoclonal antibody and tag-linked antigen, Journal of General Virology (1992), 73, 653-660

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