General description
ANTI-FLAG® M1 Agarose Affinity Gel is a purified mouse IgG2B monoclonal antibody covalently attached to agarose.
Specificity
Binding specificity: Free N-Terminus of FLAG sequence
N-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-C
Application
For purification of N-terminal FLAG fusion proteins. Since binding is Ca2+-dependent, proteins can be eluted with a buffer containing EDTA, as well as by the standard methods using either FLAG peptide or glycine-HCl buffer, pH 3. ANTI-FLAG M1 does not bind to Met-FLAG fusion proteins, so this resin is not appropriate for purifying unprocessed, cytoplasmically expressed fusion proteins.
Affinity gel is for calcium mediated purification of N-terminal FLAG fusion proteins.
immunoprecipitation (IP): suitable
Elution - FLAG peptide, Glycine, pH 3.5 EDTA
Browse additional application references in our FLAG® Literature portal. Features and Benefits
- Typically purify fusion proteins from crude lysates to single band purity in just one chromatography step.
- Fusion protein may be eluted from affinity resin by mild elution with EDTA.
- A solution of FLAG peptide can be used for gentle, non-denaturing elution of FLAG fusion proteins.
Physical form
Suspension of beaded agarose in 50% glycerol containing 10 mM sodium phosphate, 150 mM NaCl, pH 7.4, 0.02% (w/v) sodium azide
Other Notes
ANTI-FLAG® M1 does not bind to Met-FLAG fusion proteins, so this resin is not appropriate for purifying unprocessed, cytoplasmically expressed fusion proteins.
Legal Information
ANTI-FLAG is a registered trademark of Sigma-Aldrich Co. LLC
FLAG is a registered trademark of Sigma-Aldrich Co. LLC