A Myc-tag is a polypeptide protein tag derived from the c-myc gene product that can be fused to either the C- or N-terminus of a protein through recombinant DNA technology. It can facilitate the affinity purification of the tagged protein, as well as the isolation and detection of the tagged protein.
The peptide sequence of the myc-tag is: EQKLISEEDL (or -Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-).
The Myc-tag contains 10 amino acids, and the molecular weight of the Myc-tag (EQKLISEEDL-tag) peptide is 1202 Da.
Adding a Myc-tag to a protein's C- or N-terminus can facilitate its detection, isolation, and purification by Myc-tag specific antibodies. This is particularly helpful for proteins without a detection antibody or with unknown cellular location. Sino Biological has developed 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.
Myc-tagged recombinant protein can be affinity purified directly from a cell culture lysate or supernatant. The Myc-tagged protein binds to the Myc-tag specific monoclonal antibody conjugated on an agarose gel. After washing away residual impurities, bound Myc-tag proteins can be eluted off the affinity column by high concentration of the Myc-tag peptide or by low pH buffer.
In some applications, it is desirable to remove the Myc-tag, for example, for protein crystallization. To allow cleavage of the Myc-tag, a protease cleavage site needs to be engineered between the tag and the protein. An EK cleavage site behind the Myc-tag (Myc-EK site-protein structure) can allow complete removal of the Myc-tag and the cleavage site, leaving no additional amino acids after the specific cleavage of the Myc-tag. For more information on the cleavage site and tag removal by EK and HRV-3C protease, please refer to: Enterokinase (EK), HRV-3C (human rhinovirus protease).
|Full services||From gene cloning/synthesis to purified proteins|
|Host Cells||Choices of expression hosts:
Transient transfection in HEK293 cells
Transient transfection in CHO cells
Stable expression in CHO cells
Yeast cell expression system
Insect cell/baculovirus expression system
E. coli. microbial expression system
|Myc Tag Design||Myc tag (EQKLISEEDL-tag) at N- or C-terminus|
|Cleavage Site||Option to design a cleavage site between the tag and protein
Selection of cleavage protease enzyme: Enterokinase (EK), HRV-3C (human rhinovirus protease), TEV
|Purification||Myc-tag (EQKLISEEDL-tag) specific antibody conjugated affinity clumn
Myc-tag Affinity Purification Resin
|Deliverable||Purified recombinant proteins (quantities to be specified by customer)|
|Quality Specification||typically >90% purity by SDS page|
|Typical Timeline||8-12 weeks|
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. The Myc-tag (EQKLISEEDL-tag) also can be used for detection in western blot by using a Myc-tag-specific antibody. Myc-tag can be added to either C- or N-terminus of a protein for expression in virtually all expression systems.
Sino Biological has successfully produced many proteins with the Myc-tag and purified with Myc-tag affinity chromatography. We are highly experienced in designing the Myc-tag according to the protein structure and handle any problems arising from protein purification process. Sino Biological has also developed highly sensitive Myc-tag (EQKLISEEDL-tag) specific antibodies for detection in western blot, ELISA assay, Immunofluorescence, and Immunoprecipitation. We have also developed our own Myc-tag (EQKLISEEDL-tag) affinity purification resin. For more information on our Myc-tag (EQKLISEEDL-tag) epitope antibodies, please refer to: Tag antibodies. The Myc-tag antibodies can be purchased directly from our website.
Our track-records in the past years on the successful production of about 6,000 recombinant protein bulks and over 12,000 antibodies have attracted top-10 multi-national pharmaceutical customers as well as small biotech companies worldwide through word-of-mouth, as we have not done much business development or advertisement in the past. In addition, we are starting to promote our protein production services to all research institutions and companies worldwide, as we believe more customers can benefit greatly from our capacity and experience.
If you are interested in using our protein production services to support your antibody discovery and development program, please contact us by leaving an on-line message or send us your detailed request to CROfirstname.lastname@example.org. Based on the quantity and quality requirement of your request, a formal quote will be send back to you for your review, comment, and/or acceptance.