Biomolecules can be immobilized by binding them to a support under conditions that will then determine the characteristics of the complex. Immobilization is a technique that binds a ligand (enzyme, antibody, affinity proteins like Protein A or Protein G) to a support, giving high stability and making it easier to reuse the immobilized molecule.
Glyoxal beads allow a covalent binding of agarose to lysine amino groups (similar orientation to CNBr). Glyoxal agarose beads are ready to use and require no activation. The resin has a high amount of aldehydes groups (15-25 moles Glyoxal / ml gel) providing a very good option to conjugate affinity ligands improving the stability of the ligand.
The choice of Glyoxal or Aminoethyl will depend on the biomolecule to be immobilized, the accessibility of the reactive groups, and the direction/ orientation required for the binding to the support.
Glyoxal 6% agarose beads are suitable for use with FPLC, but with pressures no more than 20 kPa. Too much pressure or too fast of a flow rate will result in diminished performance.
Choosing High and Low Density Agarose Beads
High Density Resin
Low Density Resin
High Crosslinking
Low Crosslinking
Best for smaller proteins
Best for larger proteins (e.g. antibodies)
Best for repeated use or FPLC systems
Best for batch incubations or gravity flow
Small pores
Large pores
Stronger covalent binding of ligands improves ligand stability
Weaker covalent binding improves ligand mobility after immobilization
