PureCube Phenyl Agarose

Order number: 39103

€76.00*

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Description

Hydrophobic interaction chromatography (HIC) is a reliable method to separate proteins and other biomolecules based on their hydrophobicity. Our PureCube Phenyl agarose resin is a suspension of purification beads for such an approach. If you use HIC with your sample for the first time we recommend testing all of our HIC resins before.
These include:
Feature
Usage As a matriy for Hydrophobic Interaction Chromatography (HIC)
Specificity Hydrophobic biomolecules and proteins
Bead size 100 μm
Bead Ligand Phenyl groups
Phenyl group density 40 µmol/ml
BSA binding capacity 25 mg/ml
lysozyme binding capacity 45 mg/ml
pH stability 2-13
Filling quantity Delivered as a 50 % suspension
Matrix 7.5% cross-linked agarose
Short Term Storage In neutral buffer at 4°C
Long Term Storage 20 mM sodium acetate, 20% ethanol, pH 6.5 at 4 °C
Chemical Structure of Phenyl ligand Phenyl group structure

Lab Results

Purification Resins
Hydrophobic interaction chromatography is a well-established method to purify proteins according to their surface charge and hydrophobicity. Chromatography matrices that provide a hydrophobic surface are used for separation. This method does not require any affinity tags to be fused to the protein to be purified. Therefore it is suitable for the purification of both native and recombinant proteins. We offer hydrophobic interaction matrices, with phenyl, octyl, or butyl groups on the surface. For a novel protein, comparing different matrices is strongly recommended, and a HIC screening set with small quantities of each matrix is available. Our PureCube Agaroses are based on BioWorks Workbeads, providing highest binding capacities and reproducible results, especially in FPLC chromatography applications.

Binding
Proteins might precipitate at very high salt concentrations, therefore it is recommended to test in small scale experiments if the protein of interest stays in solution at the salt concentration required for binding.

Elution
Bound proteins can be eluted with low-salt buffers. It is highly recommended to use a salt gradient (e.g. ammonium sulfate or sodium chloride) for elution to ensure that the protein is kept in the optimal salt concentration required for stability.
Phenyl Agarose resin by Cube Biotech in use for HIC
Figure 1: Purification of two test proteins on PureCube Phenyl Agarose and a leading competitor product. For the larger test protein (BSA, 66 kDa) yields of about 25 mg/mL resin were obtained with both HIC resins. For the smaller test protein (lysozyme, 14 kDa) yields were about 4x higher on the PureCube matrix (PureCube: 45 mg/mL, competitor G: 11 mg/mL). Purification was done in batch, with binding and washing at 1.5 M ammonium sulfate, 100 mM NaH2PO4, pH 7.0, and elution at 100 mM NaH2PO4, pH 7.0. E1, E2: elution fractions 1 and 2.

Video

Video Guide - Hydrophobic Interaction Chromatography (Jump to 8:04)
Video Guide - How to pack FPLC cartridges

FAQ

Can I get the datasheet for the PureCube Phenyl Agarose?

What can I do with these beads?

The beads are meant for Hydrophobic Interaction Chromatography or HIC for short. This purification method separates proteins and other biomolecules based on their hydrophobicity. How the process works in detail can be found in THIS VIDEO GUIDE