Phenyl Agarose HP
Based on 1 Customer Validation
MCE Phenyl Agarose HP is a high-resolution hydrophobic chromatography medium formed by covalently coupling phenyl ligands to agarose. It is suitable for laboratory-scale and industrial-scale purification of biomolecules.
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Storage :
4°C, 5 years
Do not dry or freeze
- Manual (3240KB)
- COA
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
Hydrophobic Interaction Chromatography (HIC) media are designed to purify biomolecules based on differences in surface hydrophobicity. Common ligands such as butyl or phenyl are covalently attached to agarose or synthetic resins. Proteins bind in high-salt conditions and are gently eluted by decreasing salt concentration, preserving structure and activity. HIC is widely used in downstream processing of peptides, antibodies, and protein therapeutics, from lab-scale to industrial applications.
MCE Phenyl Agarose HP is a high-resolution hydrophobic chromatography medium formed by covalently coupling phenyl ligands to agarose. It features high mechanical strength, fast flow rate, strong selectivity, high recovery of biological activity, and environmental safety. The medium is resistant to strong acids and bases, allowing for in-place cleaning (CIP) and high-temperature sterilization. Compared to reverse-phase chromatography media, it has a lower ligand density and milder elution conditions, which better preserve the biological activity of target molecules. It is suitable for laboratory-scale and industrial-scale purification of biomolecules.
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 PB, 1.0-2.0 M ammonium sulfate, pH 7.0 |
| Elution Buffer | 20 mM PB, pH 7.0 |
| Storage Buffer | 20% ethanol |
Notes: a. Prepare all buffers using ultrapure water and sterilize them by filtration through a 0.45 μm or 0.22 μm membrane;
b. The formulations are provided for reference. Optimize the ammonium sulfate concentration according to the hydrophobicity of the target protein;
c. The general principle of hydrophobic interaction chromatography is sample loading under high-salt conditions and elution by decreasing the salt concentration.
Sample Preparation
1. Sample Filtration: Before purification, centrifuge the sample or filter it through a 0.22 μm or 0.45 μm membrane.
2. Condition Adjustment: Dilute, dialyze, or exchange the sample into binding/wash buffer so that its pH and salt concentration match those of the equilibration buffer. When adding ammonium sulfate, add it slowly with thorough mixing to avoid protein precipitation.
Protein Purification
1. Column Packing: Pack Phenyl Agarose HP into a suitable chromatography column and connect the column to the chromatography system.
2. Column Equilibration: Equilibrate the column with 5 column volumes of binding/wash buffer. Repeat the procedure 2-3 times.
3. Sample Loading: Load the sample using a pump or sample loop and collect the flow-through.
Notes: a. Select an appropriate resin volume according to the protein amount and do not exceed the binding capacity of the column;
b. High sample viscosity or large volumes may increase column backpressure.
4. Washing: Wash the column with 10-20 column volumes of binding/wash buffer to remove unbound or weakly bound contaminating proteins. Collect the wash fractions until the UV absorbance baseline stabilizes.
5. Elution: Elute with 5-10 column volumes of elution buffer using step elution, or apply a linear decreasing ammonium sulfate gradient. Collect the eluate in separate fractions.
6. Regeneration: Wash the column thoroughly with 5-10 column volumes of elution buffer, followed by 10-20 column volumes of ddH2O.
Optional: Cleaning-in-Place (CIP)
1. Removal of Precipitates and Tightly Bound Contaminants: Wash with 3-5 column volumes of 1 M NaOH, followed by 10-20 column volumes of ddH2O.
2. Removal of Lipid and Strongly Hydrophobic Contaminants: Wash with 3-5 column volumes of 70% ethanol or 30% isopropanol, followed by 10-20 column volumes of ddH2O.
Storage
After final cleaning, equilibrate the column with 5-10 column volumes of storage buffer, disconnect the column, and store at 4°C.
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 | HY-K0270-5 mL | HY-K0270-25 mL | HY-K0270-100 mL | HY-K0270-500 mL |
|---|---|---|---|---|
| Phenyl Agarose HP | 5 mL | 25 mL | 100 mL | 500 mL |
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