Thermo Scientific Pierce Pierce NHS-Activated Agarose Spin Columns, 0.2 mL contain a high-quality, amine-reactive, beaded-agarose resin for rapid and stable immobilization of proteins, peptides and other ligands via primary amines.

Features of NHS-Activated Agarose:

• Easy to use—immobilize in a simple one-step reaction with minimal hands-on time
• Rapid and efficient—greater than 85% coupling for most proteins within 30 minutes
• Innovative format—the dry agarose concentrates the sample, making it ideal for immobilizing dilute proteins
• Safe—No hazardous chemicals needed (e.g., sodium cyanoborohydride, cyanogen bromide)
• Versatile—affinity resin is adaptable to column and batch affinity chromatography techniques and FPLC applications
• Compatible—use with any primary amine-containing compound
• Reusable—the leak-resistant chemistry means you can reuse the affinity resin
• High binding capacity—coupling capacity of >25 mg/mL (dry resin)

NHS-activated agarose is crosslinked, 6% beaded agarose resin that contains N-hydroxysuccinimide (NHS) functional groups. The activated resin reacts with primary amines to form stable amide linkages that covalently immobilize antibodies or other proteins for use in affinity purification procedures. Pierce NHS-Activated Agarose has a coupling capacity greater than 25 mg/mL for the reconstituted dry resin.

Applications:
• Rapid Immobilization of goat anti-mouse and anti-rabbit antibodies in order to purify IgG produced in animals or hybridomas.
• Bulk immobilization of Protein A for purification of monoclonal antibodies
• Immobilization of ligands for purification of recombinant proteins

Pierce NHS-Activated Agarose resin uses reliable NHS-ester Chemistry and does not require hazardous chemicals for immobilization. Other amine-reactive supports, such as periodate-oxidized resins, use toxic sodium cyanoborohydride to stabilize the reaction linkage to primary amines and take 4 to 6 hours to complete. Traditional methods such as cyanogen bromide-activated supports also couple amines; however, this chemistry results in nonspecific binding and constant slow leakage of the coupled ligand. Reactions with Pierce NHS-Activated Agarose are complete in less than one hour and yield much more stable linkages.

The NHS-Activated Agarose coupling reaction is performed in an amine-free buffer at pH 7-9. Protein coupling efficiency is typically greater than 80%, regardless of the ligand's molecular weight or pI. Once a ligand is immobilized, the prepared resin can be used for multiple affinity purification procedures. The crosslinked beaded agarose has fast linear flow potential, making it useful for gravity-flow and low- to medium-pressure applications.

The unique dry form does not require storage in or removal and disposal of the acetone solvent. In addition, the dry resin is ideal for coupling reactions with dilute samples because it concentrates the sample as the resin swells, reducing the volume of the starting material and resulting in highly efficient ligand immobilization.

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