c-Myc tag properties and uses:
c-Myc tags are decapeptide tags derived from the c-myc gene product that can be encoded within DNA sequences and be positioned at either end of the protein of interest. C-Myc tags can be used for affinity purification, co-precipitation and also as epitope tags for detection by western blotting.
Anti-c-Myc antibodies are efficient for detection in a range of formats. The c-Myc tag sequence, from N to C: N-Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-C. It has a molecular weight of 1202 Da.
Immunoprecipitation of proteins using c-Myc tags: the principle
A typical c-Myc immunoprecipitation is as follows:
- Harvest and lyse host cells with expressed c-Myc-tagged protein, and clarify by centrifugation
- Incubate the lysate with anti-a-Myc antibodies and a slurry of Protein A-capture gel beads, centrifuge and wash.
- Elute the bound c-Myc-tagged protein with one of (a) a low pH glycine buffer, (b) SDS, (c) high pH urea buffer.
Our product range
Oxford Genetics provides a broad range of plasmids containing N-terminal and C-terminal c-Myc tags. In some plasmids the tags flank reporter genes, while in others they are positioned adjacent the MCS enabling you to insert your own gene in frame as required. The plasmid structure shown contains the c-Myc tag upstream of the MCS (where you can insert your gene of interest under the CMV promoter) and provides a thrombin cleavage site, allowing you to remove the c-Myc tag after production of your protein if you wish. It also contains the puromycin resistance gene under control of the ubiquitin promoter, for selection in mammalian cells, and kanamycin for selection in bacterial cells.
Our Plasmid Builder facility also provides a simple means for you to design an efficient strategy for any modifications or cloning manipulations to these plasmids. Finally, as always, although we have designed these plasmids for simple and efficient cloning, we are happy to undertake any cloning steps that you prefer to outsource. You can access all these options through the Plasmid Builder button below.
Simple plasmids containing c-Myc tags: