MBP Agarose (Dextrin) 6FF
Based on 1 Customer Validation
MCE MBP Agarose (Dextrin) 6FF is prepared by covalently coupling dextrin to an agarose matrix. It features high binding capacity, excellent specificity, and superior ligand stability. It can achieve one-step purification of MBP fusion protein.
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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 MBP Agarose (Dextrin) 6FF is prepared by covalently coupling dextrin to an agarose matrix. It features high binding capacity, excellent specificity, and superior ligand stability. This resin enables one-step purification of MBP fusion proteins. The bound fusion protein can be gently eluted using 10 mM maltose, which helps preserve the biological activity of the target protein. The MBP tag can be subsequently removed by site-specific protease cleavage.
Each 1 mL of total volume contains 0.5 mL of resin. Thoroughly resuspend the resin before use and mix well before aliquoting.
The specifications of the product correspond to the actual resin volume, with the resin content of 50%.
Protocol
Recommended Buffers (Not Provided)
| Buffer | Composition |
|---|---|
| Binding/Wash Buffer | 20 mM Tris-HCl, 200 mM NaCl, 1 mM EDTA, pH 7.4 |
| Elution Buffer | 20 mM Tris-HCl, 200 mM NaCl, 1 mM EDTA, 10 mM maltose, pH 7.4 |
| Storage Buffer | 1× PBS containing 20% ethanol |
Note: Prepare buffers with high-purity water and filter them through a 0.22 μm or 0.45 μm membrane. Process samples under cold conditions and clarify them thoroughly.
Sample Preparation
Resuspend cells or bacterial pellets expressing the MBP fusion protein in binding/wash buffer, lyse under cold conditions, and centrifuge to remove insoluble material. Filter through a 0.22 μm or 0.45 μm membrane before loading. Dialyze or desalt samples that are incompatible with the binding conditions.
Column Method
1. Column Packing and Equilibration: Thoroughly resuspend the MBP Agarose (Dextrin) 6FF, pack it into a suitable column, and equilibrate with at least 5 column volumes of binding/wash buffer.
2. Sample Loading: Load slowly using a pump, sample loop, or gravity flow and collect the flow-through. Repeated loading may improve binding efficiency.
3. Washing: Wash with 10-15 column volumes of binding/wash buffer until the OD280 or UV absorbance reaches a stable baseline.
4. Elution: Elute with 3-5 column volumes of elution buffer and collect the MBP fusion protein in fractions.
5. Regeneration and Storage: Wash thoroughly with binding/wash buffer and ddH2O, equilibrate with storage buffer, and store at 2-8°C.
Centrifugation Method
1. Resin Pretreatment: Transfer the required amount of resin suspension into a centrifuge tube, centrifuge and discard the storage solution, and wash with 5 resin volumes of binding/wash buffer 2-3 times.
2. Binding: Add the clarified sample and incubate on a rotator at 4°C for 1-2 h or overnight.
3. Washing: Centrifuge, discard the supernatant, and wash with 5 resin volumes of binding/wash buffer 3-5 times.
4. Elution: Add 3-5 resin volumes of elution buffer, incubate at room temperature for 5-10 min, centrifuge, and collect the supernatant. Repeat 2-3 times if necessary.
Storage
4°C, 2 years
Do not dry or freeze
Attention
1. Ensure the resin is thoroughly resuspended before use.
2. It is recommended to replace the storage buffer every 2-3 months to prevent microbial contamination due to ethanol evaporation.
3. To minimize protein degradation, protease inhibitor cocktails (MCE Cat. No. HY-K0010, HY-K0011) are highly recommended.
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 | HY-K0252-1 mL | HY-K0252-5 mL | HY-K0252-25 mL | HY-K0252-100 mL |
|---|---|---|---|---|
| MBP Agarose (Dextrin) 6FF | 1 mL | 5 mL | 25 mL | 100 mL |
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