Advantages
• Time saving: save 15-30 min comparing with agarose beads.
• Simple operation: Magnetic separation and centrifugation-free.
• High protein loading capacity.
• High specificity.
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Description
Anti-Flag magnetic beads is based on hydroxyl magnetic beads covalently coupling with high quality mouse IgG1 monoclonal antibody. With high loading of Flag-tagged protein (more than 1.1 mg protein/mL) and high specificity, it is recommended to use for co-immunoprecipitation and protein purification.
Properties
| Antibody isotype | Mouse IgG1 |
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| Antibody purification | Protein A purified |
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| Application | Immunoprecipitation and protein Purification |
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| Recommended volume | IP: 10 μl gel for 500 μl crude protein solution |
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| Binding capacity | Minimum 1.1 mg protein eluted per ml of magnetic beads |
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| Binding characteristics | Met-N-terminal FLAG fusion protein: Met-FLAG–Protein
N-terminal FLAG fusion protein: FLAG–Protein
C-terminal FLAG fusion protein: Protein-FLAG |
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| Shipped in | Blue ice |
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Storage
Store at 2-8°C for 2 years. DO NOT freeze or centrifuge Magnetic Beads.
Protocol
Trouble Shooting
| Problem | Possible Reason | Suggested Improvement |
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| High background | Nonspecifically binding of proteins to the antibody, megnetic beads or EP tubes | Pre-clear lysate to remove nonspecific binding proteins.
After suspending beads for the final wash, transfer the entire sample to a clean EP tube and then centrifugation. |
| Washing times are not sufficient. | Increase the number of washes.
Increase duration time of washes. |
| No signal is observed. | Flag tagged protein is not expressed in the sample. | Make sure the protein of interest contains the FLAG sequence.
Prepare the fresh lysate.
Use appropriate protease inhibitors. |
| Incubation times are inadequate. | Increase the incubation times. |
| Interfering substance is present in sample. | The lysate may contain high concentrations of dithiothreitol (DTT), 2-mercaptoethanol, or other reducing agents.
Excessive detergent concentration may interfere with the antibody-antigen interaction. |
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Anti-Flag magnetic beads
