Carboxyl-activated agarose beads can be used for the immobilization of amine containing molecules. The coupling chemistry is carbodiimide-based and is a successful and well-documented technique.
Carboxyl-activated SepFast forms chemically stable amide linkages with ligands containing primary amine groups. It has a long spacer (9 atoms). This pre-activated agarose base matrix can be readily employed to make various custom affinity chromatography media for both small scale and large scale purification applications.
Product Properties:
Carboxyl activated SepFast 4HF is made of highly cross-linked 4% beaded agarose. It shows high mechanical rigidity allowing high flow throughput with reduced back pressure.
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 large protein molecules.
The base matrix is activated by attaching a long hydrophilic spacer arm to the backbone. It reacts directly with the primary amine groups in molecules to be immobilized in the presence of a carbodiimide agent. Carboxyl-activated SepFast is supplied as an aqueous suspension in 20% ethanol.
Group to be coupled
-NH2
Matrix
Highly cross-linked 4% agarose beads (for 4HF)
Particle size
50 – 150 µm
Spacer arm
9 atoms
Activation level
8 - 19 µmol carboxyl / ml medium
pH stability
2 -13 (ligand dependent)
Chemical stability
Compatible with all commonly used aqueous chemicals, provided the ligand to be coupled can withstand
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