V5 Tag: Properties
V5 Tag: Properties, origin, specifications, antibodies & Nanobodies
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.
- 3D structure of V5 tag
- Origin
- Properties
- Size of V5 tag
- Specifications of the V5 tag
- V5 epitope tag sequences
- Applications
- V5 tag vs. Flag tag
- V5 tag vs. HA tag
- How to elute V5 tag?
- V5 tag plasmids
- V5 tag antibodies and Nanobodies - Test sample request
- V5 tag beads
- References
1. 3D structure of V5 tag
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 |
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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 |
![]() |
![]() |
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.
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-Trap Magnetic Particles M-270: anti-V5 Nanobody conjugated to magnetic particles
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