Blue Agarose 6FF
Based on 1 Customer Validation
MCE Blue Agarose 6FF can be used for the purification of biological macromolecules such as albumin, interferons, nucleotide-dependent enzymes (e.g., kinases, dehydrogenases), α2-macroglobulin, and coagulation factors.
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Storage :
4°C, 5 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
MCE Blue Agarose 6FF is an affinity chromatography medium in which Cibacron Blue 3G-A is covalently coupled to agarose.
MCE Blue Agarose 6FF is an affinity chromatography medium in which Cibacron Blue 3G-A is covalently coupled to agarose.
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 |
|---|---|
| Equilibration/Wash Buffer | 20-50 mM Tris-HCl, pH 7.0-8.0 |
| High-Salt Elution Buffer | Equilibration buffer containing 1-2 M NaCl |
| Specific Elution Buffer | An appropriate substrate, cofactor, or competitive ligand selected for the target protein |
| 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. Optimize the pH, salt concentration, and elution conditions according to the target molecule.
Sample Preparation
Clarify and filter the sample. Adjust its pH and ionic strength close to the equilibration buffer by dialysis, desalting, or dilution when necessary.
Column Method
1. Column Packing and Equilibration: Thoroughly resuspend the Blue Agarose 6FF, pack it into a suitable column, and equilibrate with at least 5 column volumes of equilibration/wash buffer.
2. Sample Loading: Load slowly using a pump, sample loop, or gravity flow and collect the flow-through. Do not exceed the binding capacity of the medium.
3. Washing: Wash with 5-10 column volumes of equilibration/wash buffer until the OD280 or UV absorbance reaches a stable baseline.
4. Elution: Elute nonspecifically with buffer containing 1-2 M NaCl, or use a substrate, cofactor, or competitive ligand selected according to the target protein. Collect fractions and identify the target fractions by protein analysis.
5. Regeneration: Remove residual bound material with a high-salt buffer, then restore the initial conditions using ddH2O and equilibration buffer. If compatible with the medium, mild alkaline cleaning may be used when required.
6. Storage: Equilibrate with storage buffer and store at 2-8°C. Do not freeze or dry.
Centrifugation Method
1. Resin Pretreatment: Transfer an appropriate amount of resin suspension into a centrifuge tube, centrifuge and discard the storage solution, and wash with 5 resin volumes of equilibration/wash buffer 2-3 times.
2. Binding: Add the prepared 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 equilibration/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, 5 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. This product is for R&D use only, not for drug, household, or other uses.
4. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K0254-5 mL | HY-K0254-25 mL | HY-K0254-50 mL | HY-K0254-100 mL |
|---|---|---|---|---|
| Blue Agarose 6FF | 5 mL | 25 mL | 50 mL | 100 mL |
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