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Description
| Application | For purification of N-terminal FLAG fusion proteins. Binding is Ca2+-dependent, so 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. |
| Biochem/physiol Actions | Binds to the FLAG epitope when it is located at the free amino-terminus of a fusion protein. Does not bind to Met-FLAG fusion proteins, so will not recognize unprocessed, cytoplasmically expressed proteins. Binding is Ca2+-dependent; the complex dissociates in the absence of calcium ions. |
| 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 in 10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% (w/v) sodium azide |
| Legal Information | ANTI-FLAG is a registered trademark of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co. |
Properties
| clone | M1 (monoclonal) |
| form | buffered aqueous suspension |
| isotype | IgG2b |
| capacity | ≥0.6 mg/mL, resin binding capacity (FLAG-BAP) |
| shipped in | wet ice |
| storage temp. | 2-8°C |
Safety
