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The c-Myc tag is derived from the c-myc gene product, and this polypeptide protein tag can be fused to either the N- or C-terminus of a target protein based on recombinant DNA technology. The c-Myc tag facilitates isolation, purification, and detection of the recombinant tagged protein. The sequence of the c-myc tag is -Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-.
c-Myc-tagged protein expression system
Fusion tag: c-Myc tag.
Purification method: c-Myc tag affinity chromatography.
Tag removal: Protease such as EK and HRV-3C.
Products: Purified recombinant proteins.
Advantages of c-Myc tag
1. c-Myc tag is very small (only 11 amino acids long), therefore, it is able to minimize effect on properties of target protein (conformation, function and localization).
2. c-Myc tag is able to allow normal cell physiology and increase protein expression level since it provides a low metabolic load.
3. c-Myc tag helps in recombinant protein expression monitoring in a wide variety of expression systems.
4. c-Myc tag is very useful in co-immunoprecipitation studies, Western blots, and flow cytometry.
5. c-Myc tag helps in subcellular localization monitoring of target proteins in mammalian cells.
6. c-Myc tag can be added to either the C- or N-terminus of target protein, facilitating the detection, isolation, and purification of recombinant protein.
It is useful to displace c-Myc-tagged proteins with c-Myc peptide to specifically bind anti-c-Myc antibodies in immunoassays. c-Myc peptide is considered to regulate the transcription process of growth-related gene. Sequence of c-Myc peptide corresponds to the 410-419 AA of the human c-myc.
c-Myc-tagged protein purification
c-Myc-tagged proteins can be directly purified from the lysate or supernatant of cell culture based on affinity chromatography. The c-Myc-tagged proteins specifically bind to the anti-Myc-tag mAb that is conjugated on an agarose gel. After washing steps to get rid of the residual impurities, you can elute the bound c-Myc tag proteins off the affinity column with low pH buffer or high concentration of the c-Myc tag peptide (0.5 mg/mL in PBS) under native conditions.
Cleavage of the c-Myc-tagged protein after purification
In some cases, c-Myc tag does not needed to be removed since it is too small to interfere the function of target protein. While in some other cases, it is preferred to perform c-Myc tag removal for better protein crystallization. A protease cleavage site is required to be inserted between the target protein and the tag in order to allow cleavage of the c-Myc tag. An EK cleavage site can be fused behind the c-Myc tag to form a c-Myc-EK site-target protein structure, which allows a specific and complete c-Myc tag removal and does not leave additional amino acids. Enterokinase (EK) and HRV-3C (human rhinovirus protease) are the most applied protease for cleavage of the c-Myc-tagged proteins.
Creative Biostructure have successfully completed numerous projects of c-Myc-tagged protein purification, and we have developed this resourceful technical support platform to help you with custom requirements. Please see the related services for more information and feel free to Contact Us.