- Matchmaker Gold yeast two-hybrid systems
- Capturem rapid purification technology
- Mass spectrometry digestion reagents
- His-tag purification
- Antibody purification
- Other tag purification
- Phosphoprotein and glycoprotein purification
- Expression systems
FLAG-tagged protein purification overview
Different technologies for FLAG-tagged protein purification: agarose and magnetic beads
The FLAG (DYKDDDDK epitope) tag is a small, widely used, well-characterized hydrophilic peptide tag (8 amino acids long). It is bound by anti-DYKDDDDK antibodies, can be incorporated on either the N- or C-terminus of target proteins, and can be expressed in bacterial, yeast, mammalian, and baculovirus-infected insect cells.
The FLAG tag is well-suited for the following applications:
- When a small tag is required to minimize its effect on protein function
- When a hydrophilic tag is required
- When a low metabolic load is needed to maintain normal cell physiology and/or to increase protein expression
- When highly specific detection is necessary
- To monitor recombinant protein expression in bacterial, yeast, mammalian, or insect cells
- For co-immunoprecipitation studies, Western blots, and flow cytometry
- To monitor subcellular localization of tagged proteins in mammalian cells
Purifying FLAG-tagged proteins using immunoprecipitation
Immunoprecipitation (IP) is a technique for separating a protein of interest from a solution (such as a cell lysate) containing thousands of different proteins by precipitating it from the solution using an antibody that binds specifically to that protein. Proteins that contain peptide tags, such as the FLAG (DYKDDDDK epitope) tag, can be isolated and concentrated using an antibody specific for this tag, provided that the antibody is attached to a solid substrate. A tag-specific antibody can also be used to precipitate protein complexes, in a process known as co-immunoprecipitation (Co-IP). We offer a choice of anti-DYKDDDDK antibodies attached to two different substrates, agarose beads or magnetic beads, for your IP and co-IP experiments.
The Magnetic DYKDDDDK Immunoprecipitation Kit offers a simple, rapid method for purifying or immunoprecipitating FLAG-tagged fusion proteins from cell lysates using magnetic beads cross-linked to a monoclonal anti-DYKDDDDK antibody (which recognizes the FLAG epitope). The kit also includes the Magnetic Beads Immunoprecipitation Buffer Set (optimized to provide extremely low background) and the ProteoGuard Protease Inhibitor Cocktail (specially designed to inhibit endogenous proteases released during cell lysis).
Anti-DYKDDDDK Beads consist of a monoclonal anti-DYKDDDDK antibody (which recognizes the FLAG epitope), crosslinked to agarose beads. These beads are used to purify or immunoprecipitate DYKDDDDK-tagged or FLAG-tagged fusion proteins from cell lysates. Our Anti-DYKDDDDK Beads and Immunoprecipitation Buffer Set can be purchased separately to make a complete co-IP kit for purification of FLAG-tagged proteins.
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