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Myc-tag: An epitope tag for protein characterization, protein interaction analysis, and purification.

Posted by Christoph Eckert on Mar 17, 2021 11:47:58 AM

Myc-tag is a peptide tag derived from the c-Myc protein. The Myc-tag can be used for many capture and detection applications such as immunoprecipitation, immunofluorescence and protein purification.

1. What is Myc- tag?
2. How does Myc-tag work?
3. 3D structure of Myc-tag
4. Origin
5. Properties
6. Size of Myc-tag
7. Specifications of the Myc-tag
8. Myc-tag epitope tag sequences
9. Applications
10. Myc-tag vs. Flag
11. Myc-tag vs. HA
12. Myc-tag vs. Spot-Tag
13. How to elute Myc-tagged proteins?
14. Myc-tagged plasmids
15. Myc-tag Nanobodies and antibodies
16. Myc-Trap: The best anti-Myc-tag antibody for immunoprecipitation
17. Myc beads
18. Reference

 

1. What is Myc-tag?

Myc-tag is an epitope tag derived from the c-Myc protein.

 

2. How does Myc-tag work?

A Myc-tag can be used to detect expression of recombinant proteins.
First, the Myc-tag should be genetically fused/cloned to a protein of interest (POI). After expression, the Myc-tagged protein can be captured and identified in crude biological samples. Common applications are immunoprecipitation, co-immunoprecipitation, immunofluorescence, ELISA, flow cytometry, protein purification, and Western blotting.

 

3. 3D structure of Myc-tag

Myc-Peptide

4. Origin

The Myc-tag is derived from the human c-myc oncogene and corresponds to amino acid residues 410-419 of the C-terminus of human c-Myc protein. The human c-Myc was discovered as a cellular homolog of v-myc oncogenes, which were identified through analyses of avian tumors1,2. C-Myc is a transcription factor.

 

5. Properties

The Myc-tag contains 10 amino acids (aa) and the sequence is EQKLISEEDL.

 

6. Size of the Myc-tag

Number of amino acids: 10
Molecular weight (MW): 1203.31Da

 

7. Specifications of the Myc-tag

Theoretical isoelectric point (pI): 4.00
Total number of negatively charged residues (Asp + Glu): 4
Total number of positively charged residues (Arg + Lys): 1

 

8. Myc-tag epitope tag sequences

Myc-tag amino acid sequence:
EQKLISEEDL

Myc-tag DNA sequence:
GAA CAA AAA CTC ATC TCA GAA GAG GAT CTG
Please note, the mentioned sequence is optimized for mammalian expression.

 

9. Applications

Myc-tagged proteins are used in immunoprecipitation (IP), protein purification, Western blotting, ELISA, flow cytometry, and immunofluorescence (IF).

 

10. Myc-tag vs. Flag-tag

Comparison of Myc-tag and Flag-tag

  Myc-tag Flag-tag
Origin human c-Myc artificial design
3D Structure Myc-tag Flag-tag2
Amino acid sequence EQKLISEEDL DYKDDDDK
Number of amino acids 10 8
Molecular weight (in Da) 1203.31 1012.98
Theoretical isoelectric point (pI) 4.00 3.97
Total number of negatively charged residues (Asp + Glu) 4 5
Total number of positively charged residues (Arg + Lys) 1 2

 

11. Myc-tag vs. HA-tag

Comparison of Myc-tag and HA-tag

  Myc-tag HA-tag
Origin human c-Myc Human influenza hemagglutinin (HA) is derived from the human influenza virus
3D Structure Myc-tag HA_2
Amino acid sequence EQKLISEEDL YPYDVPDYA
Number of amino acids 10 9
Molecular weight (in Da) 1203.31 1102.17
Theoretical isoelectric point (pI) 4.00 3.56
Total number of negatively charged residues (Asp + Glu) 4 2
Total number of positively charged residues (Arg + Lys) 1 0

 

12. Myc-tag vs. Spot-Tag®

Comparison of Myc-tag and Spot-Tag

  Myc-tag Spot-Tag
Origin human c-Myc Derived from human beta-catenin
3D Structure Myc-tag Spot-tag2-1
Amino acid sequence EQKLISEEDL PDRVRAVSHWSS
Number of amino acids 10 12
Molecular weight (in Da) 1203.31 1396.53
Theoretical isoelectric point (pI) 4.00 10.03
Total number of negatively charged residues (Asp + Glu) 4 1
Total number of positively charged residues (Arg + Lys) 1 2

 

13. How to elute Myc-tagged proteins?

Myc-tagged proteins are normally eluted using glycine or citric acid or by competitive elution with 1x Myc-peptide (EQKLISEEDL) or 2x Myc-peptide (EQKLISEEDLEQKLISEEDL).

 

14. Myc-tagged plasmids

Vectors for expression with Myc-tag for mammalian cells, Drosophila, etc. are available from Merck (Sigma-Aldrich) and ThermoFisher (Invitrogen).

 

15. Myc-tag Nanobodies and antibodies

For experimental analysis of Myc-tagged proteins, Nanobodies, monoclonal antibodies, and polyclonal antibodies are very useful. There is only one Nanobody available against Myc-tag and different polyclonal and monoclonal antibodies available/on the market.
•Recombinant Nanobody: Myc-Trap® for immunoprecipitation
•Monoclonal antibody: Myc-tag monoclonal antibody (9E1) for immunofluorescence (IF), immunoprecipitation (IP), ELISA, and Western blotting (WB).
•Polyclonal antibodies: various suppliers

 

16. Myc-Trap: The best anti-Myc-tag antibody for immunoprecipitation

For immunoprecipitation, ChromoTek’s Myc-Trap is recommended as Nanobodies have lower background compared to antibodies.
• No heavy & light antibody chains in downstream applications
• Efficient and fast pulldown of Myc-tagged proteins
• One-step immunoprecipitation
• Peptide elution of native proteins

Myc-Trap Agarose Immunoprecipitation-1Myc-Trap Agarose for immunoprecipitation of Myc-tagged proteins.

 

The Myc-Trap has more than 60 citations on citeAB.

Would you like to test the Myc-Trap yourself?

Request your free Myc-Trap sample here

 

 

17. Myc beads

Chromotek offers Myc beads, e.g., an anti-Myc-tag Nanobody conjugated to beads for immunoprecipitation:
Myc-Trap Agarose: anti-V5 Nanobody conjugated to agarose beads
Myc-Trap Magnetic Agarose: anti-V5 Nanobody conjugated to magnetic agarose beads

 

18. Reference

(1) Duesberg PH, Vogt PK. Avian acute leukemia viruses MC29 and MH2 share specific RNA sequences: evidence for a second class of transforming genes. Proc Natl Acad Sci U S A. 1979;76:1633–1637. [PMC free article] [PubMed]
(2) Sheiness D, Bishop JM. DNA and RNA from uninfected vertebrate cells contain nucleotide sequences related to the putative transforming gene of avian myelocytomatosis virus. J Virol. 1979;31:514–521. [PMC free article] [PubMed]

 

Disclaimer:

Only for research applications, not for diagnostic or therapeutic use!

ChromoTek, Myc-Trap and Spot-Tag are registered trademarks of ChromoTek GmbH, part of Proteintech Group. Nanobody is a registered trademark of Ablynx, a Sanofi company. Sigma-Aldrich is a company from Merck KGaA. Invitrogen™ is a part of Thermo Fisher Scientific Inc. Other suppliers’ products may be trademarks or registered trademarks of the corresponding supplier. Statements on other suppliers’ products are given according to our best knowledge.

Topics: Myc-tag, Spot-Tag, HA-Tag, Flag-Tag

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