c-Myc-tag is an epitope tag derived from human c-Myc oncoprotein and corresponds to amino acid residues 410-419 of the C-terminus of human c-Myc protein. Myc tag peptide is a synthetic peptide with a molecular weight of 1,203 Da. The peptide sequence is EQKLISEEDL (or -Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-).
The recombinant protein tagged with c-Myc tag can be recognized by a well-known high-affinity 9E10 antibody. Overexpression of c-myc has been reported in a wide variety of human cancers. The Myc tag sequence, EQKLISEEDL, is derived from the loosely defined epitope of the monoclonal antibody 9E10. Though it can be added to the C-terminus or N-terminus of a protein, it is not recommended to append the c-Myc tag directly behind the signal peptide of a secretory protein because the tag can interfere with translocation into the secretory pathway. In any case, the c-Myc tag can be used in many different assays such as subcellular localization studies by immunofluorescence or detection by western blot.
c-Myc tags for protein purification
Myc-tag (EQKLISEEDL-tag) has been widely used in recombinant protein expression and production. The Myc-tag allows simple and efficient affinity purification of the tagged protein using Myc-tag specific antibody conjugated to agarose-beads. Adding a c-Myc-tag to a protein's C- or N-terminus can facilitate its detection, isolation, and purification by c-Myc-tag antibodies. This is particularly helpful for proteins without a detection antibody or with unknown cellular location. Creative Diagnostics has developed c-Myc-tag specific antibodies for ELISA, Western Blotting, Immonofluorecence, and Immunoprecipitation detection of the Myc-tagged protein. It can also be conjugated to agarose beads for Myc-tagged protein affinity purification.
Fig 1. Analysis for protein study
Under native conditions, the elution of c-Myc-tagged proteins can be achieved by the addition of the c-Myc tag peptide (0.5 mg/mL in PBS) which competes with the recombinant proteins.
References
Hillman MC, Yang LS, Sun S, et al. A comprehensive system for protein purification and biochemical analysis based on antibodies to c-myc peptide[J]. Protein Expr Purif. November 2001.
Zhao X, Li G, Liang S. Several affinity tags commonly used in chromatographic purification[J]. J Anal Methods Chem. December 2013.
Hillman MC, Yang LS, Sun S, et al. A comprehensive system for protein purification and biochemical analysis based on antibodies to c-myc peptide. Protein Expression and Purification. 2001;23(2):359–368.
Loading ......
Fig 1. Analysis for protein study