Anti-MBP Magnetic Beads
Based on 1 Customer Validation
Anti-MBP Magnetic Beads are suitable for the detection and purification of MBP-tagged fusion proteins, as well as for immunoprecipitation (IP) and co-immunoprecipitation (Co-IP) applications. The 1 mL is defined as the base specification. All larger sizes correspond to incremental volumes of this base.
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Storage :
4°C, 2 years
Do not dry or freeze
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
Maltose Binding Protein (MBP) is a maltose/maltodextrin-binding protein derived from Escherichia coli, with a molecular weight of approximately 42 kDa.
The MBP tag offers the following advantages: it significantly enhances the solubility of heterologous proteins, particularly those expressed as inclusion bodies; it can be expressed in Escherichia coli or yeast systems to improve the expression level of target proteins; it promotes proper protein folding; the MBP tag can be removed by specific proteases such as PreScission, TEV, or Thrombin; it enables a simple and highly specific purification process. Additionally, the MBP tag is suitable for studies involving protein expression, localization, purification, and immunoassays. Due to these advantages, the MBP tag is widely used in protein research.
MCE Anti-MBP Magnetic Beads are prepared by covalently coupling high-quality mouse monoclonal anti-MBP antibodies to magnetic beads. These beads offer high binding capacity, excellent specificity, and stable performance. Anti-MBP Magnetic Beads are suitable for the detection and purification of MBP-tagged fusion proteins, as well as for immunoprecipitation (IP) and co-immunoprecipitation (Co-IP) applications.
Protocol
Recommended Buffers (Not Provided)
| Buffer | Composition |
|---|---|
| Wash Buffer | TBST: 50 mM Tris-HCl, 150 mM NaCl, 0.5% Tween-20, pH 7.4 |
| Elution Buffer | 0.1 M glycine, pH 3.0 |
| Neutralization Buffer | 1 M Tris-HCl, pH 8.0 |
Sample Preparation
Serum: If the target protein is abundant, dilute the sample to a final target-protein concentration of approximately 50-100 μg/mL and keep it on ice.
Suspension cells: Collect cells by centrifugation at 500 × g for 10 min at 4°C and wash twice with PBS. Add 5-10 μL of lysis buffer per mg of cells together with a suitable protease inhibitor, lyse on ice for 10 min, and centrifuge at 14,000 × g for 10 min at 4°C to collect the supernatant.
Adherent cells: Remove the medium, wash twice with PBS, scrape the cells, add 20-30 μL of lysis buffer per 1×105 cells together with a protease inhibitor, lyse on ice for 10 min, and centrifuge at 14,000 × g for 10 min at 4°C to collect the supernatant.
Immunoprecipitation
1. Bead Pretreatment: Thoroughly resuspend the beads. A typical reaction uses approximately 20 μL of beads per 500 μL of lysate. Wash with 500 μL of wash buffer, perform magnetic separation, and discard the supernatant. Repeat 2-3 times.
2. Sample Binding: Add the cell lysate and incubate on a rotator for 1-2 h at room temperature or for 2-4 h/overnight at 4°C. Perform magnetic separation and collect the supernatant.
3. Washing: Wash with 1 mL of wash buffer 2-3 times, performing magnetic separation after each wash.
4. Elution: For acidic elution, add 50-100 μL of elution buffer, incubate at room temperature for 5-10 min, perform magnetic separation, and immediately add 1/10 volume of neutralization buffer. For denaturing elution, add 1× SDS-PAGE Loading Buffer, heat at 95°C for 5 min, perform magnetic separation, and collect the supernatant.
Note: Acid-eluted proteins may be used for downstream functional analysis. Denatured eluates are suitable for SDS-PAGE or Western blotting.
Storage
4°C, 2 years
Do not dry or freeze
Attention
1. Do not centrifuge, dry or freeze the magnetic beads, which will cause the beads to aggregate and lose binding affinity.
2. To minimize protein degradation, protease inhibitor cocktails (MCE Cat. No. HY-K0010, HY-K0011) are highly recommended.
3. Do not use cell lysates containing dithiothreitol (DTT), as DTT may cause the antibody to detach from the magnetic beads.
4. This product is for R&D use only, not for drug, household, or other uses.
5. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K0250-1 mL | HY-K0250-5 mL |
|---|---|---|
| Anti-MBP Magnetic Beads | 1 mL | 1 mL × 5 |
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