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The FLAG Expression System is a proven method to express, purify and detect recombinant fusion proteins. FLAG and 3xFLAG are useful in Western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure, and protein localization. These small hydrophilic tags significantly improve detection and purification of recombinant fusion proteins when used with our highly specific and sensitive ANTI-FLAG® antibodies.
Technical Publications & Literature | Overview of FLAG-Protein Expression System | Vectors for Bacterial and Mammalian Systems | Comparative Sensitivity of 3xFLAG | Cytochemical Staining using FLAG | FLAG and 3xFLAG Products Technical Publications & Literature
The FLAG® system utilizes a short, hydrophilic 8-amino acid peptide (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) which is fused to the recombinant protein of interest when expressed from a pFLAG vector. The FLAG peptide includes the binding site for several highly specific ANTI-FLAG monoclonal antibodies (M1, M2, M5) and polyclonal antibodies and conjugates, each with different recognition and binding characteristics. The FLAG peptide is likely to be located on the surface of a fusion protein because of its hydrophilic nature. As a result, the FLAG peptide is more likely to be accessible to antibodies and to cleavage by enterokinase (Ek). In addition, because of the small size of the FLAG® peptide tag, it is not likely to obscure other epitopes, domains, or alter function, secretion, or transport of the fusion protein. The 3X FLAG system improves upon the original system by fusing 3 tandem FLAG epitopes to a recombinant protein (Fig. 1). A 10-20 fold increased detection enhancement has been shown using 3X FLAG fusions (Fig. 2, 3). As with the original FLAG® tag, it is hydrophilic, contains an enterokinase cleavage site and is relatively small (22 amino acids), therefore, the risk of altering protein function, blocking other epitopes or decreasing the solubility is minimized.
Vectors for Bacterial and Mammalian Systems
Comparative Sensitivity of 3X FLAG
Each protein fusion tag was cloned separately on the C-terminal of GST. Cytochemical Staining using FLAG
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