Activated Sepharose. Binds up to 20 mg protein per ml of settled resin (BSA as representative protein). Product code: AAG. Provided in acidified 90% acetone 10% acetic acid for stability. Store at -20 C in small amounts. Reacts with amines at pH 8-9. Quickly loses activation if stored in neutral or basic solutions. Quantity: Clear. A DoE approach was selected to optimize the coupling efficiency of DFE to NHS-activated cross-linked agarose resin. The absolute amount of DFE immobilized on the resin increased with the mass of DFE injected into the column and plateaued at 15 gL -1 whereas the coupling yield declined.
NHS-Activated Agarose consists of 6% 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. Features.
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 generatedApplications. Peptide coupling for antibody purification. Protein purification. Antibody purification. Purify protein interacting partners.
Sepharose is a tradename for a crosslinked, beaded-form of, a polysaccharide polymer material extracted from seaweed. Its brand name is a derived from Separation- Pharmacia-Aga rose. A common application for the material is in chromatographic separations of biomolecules.Sepharose is a registered trademark of (formerly: Pharmacia, Pharmacia LKB Biotechnology, Pharmacia Biotech, Amersham Pharmacia Biotech, and Amersham Biosciences).Various grades and chemistries of sepharose are available.Iodoacetyl functional groups can be added to selectively bind side chains and this method is often used to immobilize. Sepharose/agarose, combined with some form of activation chemistry, is also used to immobilize, and other and peptides through attachment to the resin.
Common activation chemistries include (CNBr) activation and reductive amination of to attach proteins to the agarose resin through lysine side chains.See also References.
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March 2023
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