The base matrix is activated by attaching haloacetyl group through a long hydrophilic spacer arm which facilitates much improved ligand accessibility. It reacts rapidly with molecules containing thiol groups.
Irreversible Thiol-coupling SepFast can be used for the specific immobilization of thiol containing molecules. The coupling chemistry is based on a haloacetyl activation method that is a successful and well-documented technique. The reaction to thiol groups is highly specific and formed bond is irreversible. The coupling reaction is rapid and easy to carry out. No toxic chemicals or special equipment is required.
The SepFast base matrix is highly cross-linked. It shows high mechanical rigidity allowing high flow throughput with reduced back pressure in a packed column.
Agarose has long been used for chromatographic separations due to its excellent hydrophilic and low non-specific-binding nature. The particles have an open pore structure with excellent mass transfer properties to various molecules.
Irreversible Thiol-coupling SepFast medium forms an extremely stable thioether bond with ligands containing thiol (i.e. sulphhydryl) groups. This pre-activated agarose based resin can be readily employed to make various custom affinity chromatography media for both small scale and large scale purification applications.
There is a choice of two different activated base matrices that suit ligands of various molecular weights.
- Irreversible Thiol-Coupling SepFast 4HF is made of highly cross-linked 4% beaded agarose.
- Irreversible Thiol-Coupling SepFast 6HF is made of highly cross-linked 6% beaded agarose.
Coupling manual is supplied with each product.
|Group to be coupled||-SH|
|Coupling conditions||pH 8 – 9, +4°C to +30°C, 1 - 16 h, organic solvents possible|
|Matrix||SepFast 4HF: highly cross-linked 4% agarose|
|SepFast 6HF: highly cross-linked 6% agarose|
|Particle size||50 – 150 μm|
|Coupling level||at least 3 mg BSA / ml medium|
|pH stability||3 -11 (ligand dependent)|
|Chemical stability||Stable to all commonly used aqueous chemicals, provided the ligand to be coupled can withstand|
|Storage||+4°C to +8°C|
|Irreversible Thiol-coupling SepFast 4HF||5 ml||390104|
|Irreversible Thiol-coupling SepFast 6HF||5 ml||390110|