Streptavidin-Binding Peptide Tag Agarose
Based on 1 Customer Validation
MCE Streptavidin-Binding Peptide Tag Agarose is composed of a highly cross-linked 4% agarose matrix with an engineered streptavidin mutant protein chemically immobilized onto the matrix in a highly oriented and high-density manner. It is suitable for the efficient purification of recombinant proteins carrying a streptavidin-binding peptide tag from a variety of expression systems, including E.coli, yeast, insect cells, and mammalian cells.
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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
The streptavidin-binding peptide tag consists of 8 amino acids (WSHPQFEK).
MCE Streptavidin-Binding Peptide Tag Agarose is composed of a highly cross-linked 4% agarose matrix with an engineered streptavidin mutant protein chemically immobilized onto the matrix in a highly oriented and high-density manner. It is suitable for the efficient purification of recombinant proteins carrying a streptavidin-binding peptide tag from a variety of expression systems, including E.coli, yeast, insect cells, and mammalian cells.
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 | 100 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, pH 8.0 |
| Elution Buffer | 100 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 2.5 mM desthiobiotin, pH 8.0 |
| Storage Buffer | 1× PBS containing 20% ethanol |
Note: Filter buffers and samples through a 0.22 μm or 0.45 μm membrane.
Sample Preparation
Clarify and filter soluble protein samples. If necessary, dialyze or dilute with binding/wash buffer to obtain a suitable pH and ionic strength.
Column Method
1. Column Packing and Equilibration: Thoroughly resuspend the Streptavidin-Binding Peptide Tag Agarose, pack it into a suitable column, and equilibrate with 5 column volumes of binding/wash buffer 2-3 times.
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 target protein in fractions. Use smaller fractions when a higher protein concentration is required.
5. Regeneration and Storage: Wash thoroughly with 5-10 column volumes of binding/wash buffer and deionized water, equilibrate with storage buffer, and store at 2-8°C.
Centrifugation Method
1. Resin Pretreatment: Transfer an appropriate amount of resin suspension into a centrifuge tube, centrifuge at 1,000-5,000 × g for 1 min, discard the supernatant, and wash with 5 resin volumes of binding/wash buffer 2-3 times.
2. Binding: Add the 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.
Components
| Components | HY-K0239-1 mL | HY-K0239-5 mL | HY-K0239-10 mL |
|---|---|---|---|
| Streptavidin-Binding Peptide Tag Agarose | 1 mL | 5 mL | 10 mL |
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