Epitope tags

Epitope tags serve as universal epitopes, that are being added to recombinant proteins without negatively affecting their functions. Such N-terminal or C-rerminal tags can be used as markers of the particular recombinant proteins, as tools for their purification, etc. For this purpose e.g. c-Myc protooncogen is used, as well as glutathione-S-transferase (GST) from Schistosoma japonicum, beta-Galactosidase from Escherichia coli, or horseradish peroxidase (HRP). DDDDK tag (F-tag) is popular for its small size and high hydrophilicity. Fluorescent tags, such as GFP, mCherry, mPlum, and Dendra 2 can be used for more complex studies.  

 

 
Fig. 1:  One example of possible usage of an epitope tag antibody describes detection of a recombinant protein. It is useful especially in case that there is only a limited offer of specific antibodies to this protein in the market, or in case when the recombinant protein must be distinguished from its naturally expressed variant. The antibody can be also used for isolation of the recombinant protein.
 
 
 
Table of our antibodies to epitope tags:
 

 

 


Fig. 2: Detection of transfected LST-1-c-Myc in HEK-293 cells (red) compared with nontransfected HEK-293 cells (black) using mouse monoclonal anti-c-Myc (9E10) purified, GAM-APC.
 
 

 
Fig. 3: Immunocytochemistry (confocal microscopy) of COS-7 cells transfected with expression constructs encoding fusion nuclear protein with DDDDK epitope. A - fusion nuclear protein (red) stained with purified anti-DDDDK (F-tag-01) (detection by Goat anti-mouse IgG1 Alexa Fluor® 594). B - cell nuclei stained with DAPI (blue). C - merged figures - confirmation of nuclear localization of the fusion protein; cell nuclei stained with DAPI (blue).
 
 

 
Fig. 4: Western blotting analysis of Dendra2/GPI fusion protein using rabbit polyclonal antibody PAb (836) on lysates of HEK293T/17 cells transfected with Dendra2/GPI construct; reducing and non-reducing conditions. Nitrocellulose membrane was probed with 2 µg/ml of anti-Dendra2 rabbit polyclonal antibody followed by IRDye800-conjugated anti-rabbit secondary antibody. A specific band was detected for Dendra2/GPI fusion protein at approximately 35 kDa.
 
 
 

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