Anti-c-Myc Magnetic Beads |
| Catalog No.GK30005 |
Anti-c-Myc Magnetic Beads are used for immunoprecipitation (IP) of specific c-Myc-tagged proteins expressed in bacterial and mammalian cells and in vitro expression systems.
Products are for research use only. Not for human use. We do not sell to patients.
Sample solution is provided at 25 µL, 10mM.
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Anti-c-Myc magnetic beads are used for immunoprecipitation (IP) of specific c-Myc-tagged proteins expressed in bacterial and mammalian cells and in vitro expression systems. Anti-c-Myc magnetic beads are based on amino magnetic beads, with 2 μm particle size, covalently coupling with high quality mouse IgG1 monoclonal antibody that recognizes the c-Myc-epitope tag (EQKLISEEDL) derived from the human c-Myc protein. Magnetic beads are removed from the solution manually by using a magnetic stand or automatically by using an instrument. With high loading of c-Myc-tagged protein and high specificity, Anti-c-Myc Magnetic Beads are also suitable for Co-immunoprecipitation and purification of c-Myc-tagged protein.
1. Recommended Wash Buffer
PBST: 1×PBS + 0.5% Tween-20, pH 7.4
2.Preparation of Magnetic Beads
2.1 Resuspend the Magnetic Beads in the vial (tilt and rotate for 2 minutes or gently pipette for 10 times, do not vortex). Transfer 20 μL of Anti-c-Myc Magnetic Beads suspension into a new tube.
2.2 Add 500 μL of wash buffer to the beads and gently pipette to mix. Place the tube into a magnetic stand to collect the beads against the side of the tube. Remove and discard the supernatant. Repeat this step for 2 times.
3 Protein Binding
3.1 dd 500 μL of cell lysate (the sample containing c-Myc-tagged protein) to the washed beads. For Ag binding, incubate for 2 hours at room temperature or overnight at 4ºC while gently rotating the tube.
3.2 Place the tube into a magnetic stand to collect the beads against the side of the tube. Remove and discard the supernatant.
Note: Occasional aggregation of magnetic beads during the binding process doesn’t affect experimental results.
4. Washing
Add 500 μL of wash buffer to the Magbeads-Ag complex and mix gently. Place the tube into a magnetic stand to collect the beads against the side of the tube. Remove and discard the supernatant. Repeat this step for 4 times.
5. Elution & Detection
Add 50 μL of 1× SDS-PAGE loading buffer into the tube with Magbeads-Ag complex and boil for 5 minutes. Cool and place the tube into a magnetic stand to collect the beads against the side of the tube. Keep the supernatant containing the target antigen for SDS-PAGE analysis.
Precautions：
1 The pH of Anti-c-Myc Magnetic Beads is 6-8.
2 Do not centrifuge, dry or freeze the magnetic beads. Centrifuging, drying or freezing will cause the beads to aggregate and lose binding affinity.
3 For best results, determine optimal conditions for expression of c-Myc-tagged fusion protein before attempting immunoprecipitation.
4 To minimize protein degradation, protease inhibitor cocktails (Cat.No.: GK10014) are highly recommended.
5 For the best experimental performance, it is recommended to use the magnetic stand.
6 Do not use cell lysate containing dithiothreitol (DTT). DTT may cause the c-Myc antibody to leach from the beads.
7. This product is for R&D use only, not for drug, house hold, or other uses.
Troubleshooting：
|
Problem |
Possible Cause |
Solution |
|
High background |
Nonspecifically binding of protein to the antibody, magnetic beads or EP tubes. |
Pre-clear lysate to remove nonspecific binding proteins. |
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After suspending beads for the final wash, transfer the entire sample to a clear EP tube and then magnetic separation or centrifugation. |
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Washing times were not sufficient. |
Increase the number and time of washes. |
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Little or no c-Myc-tagged protein is detected |
No or minimal tagged protein was expressed. |
Verify protein expression by SDS-PAGE or Western blot analysis of the lysate using an c-Myc-tagged positive control as a reference. |
|
Increase the amount of lysate used for IP. |
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|
Tagged protein degraded. |
Prepare fresh lysate. |
|
|
Use appropriate protease inhibitors (Cat. No.: GK10014). |
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Incubation time was inadequate. |
Prolong the incubation time. |
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Interfering substance was contained. |
Do not use cell lysate containing dithiothreitol (DTT),2-mercaptoethanol, or other reducing agents. |
|
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Excessive detergent concentration may interfere with the antibody-antigen interaction. |
| Applications |
Concentration: 10 mg/mL
Bead Diameter: 2μM Binding Capacity: >0.5 mg protein/mL beads Storage Buffer: PBS,ph 7.2 |
| Shipping | Ship with blue ice. |
| Storage Conditions | Stored at 4°C, and is stable for up to 2 years. Do not centrifuge, dry or freeze the magnetic beads. |
| Usage | For research use only! Not for use on humans. |
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
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