NHS-Activated Agarose consists of 4% cross-linked agarose that has been activated by the addition of a reactive NHS (N-hydroxysuccinimide) group. The NHS group forms covalent, chemically stable amide bonds with ligands that contain primary amines. The NHS-Activated Agarose also contains a spacer arm between the NHS group and the agarose beads, making it suitable for coupling of small proteins and peptides. The long spacer arm minimizes steric hindrance allowing high efficient binding of ligands, including small proteins and peptides.
The 4% highly cross-linked agarose beads are coupled to 6-carbon spacer arm. The terminal carboxyl group of the spacer arm is activated by esterification with the NHS group.
The coupling reaction is performed in an amine-free buffer at pH7-9 and the coupling efficiency is typically >80%, regardless of ligand’s pI or molecular weight. Once the ligand is coupled to the resin, the resin can be used for multiple affinity purifications. The resin is suitable for gravity-flow and low- to medium-pressure applications.
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- High level of activation
- Simple: One step couple reaction
- Stable: Resulting affinity column very stable, especially at pH extremes
- Spacer arm between agarose and reactive group, minimize steric hindrance
- Suitable for small proteins and peptides
- Rapid: >80% coupled in first 30 minutes
- Suitable for any ligand with primary amine
- Reusable affinity chromatography columns generated
- Peptide coupling for antibody purification
- Protein purification
- Antibody purification
- Purify protein interacting partners