Aldehyde-Activated Separopore® 4B

Aldehyde-Activated Separopore®, an amino link resin is a beaded agarose support that has been activated to contain aldehyde groups. Antibodies or other proteins with primary amines can be conjugated to the Aldehyde Separoppore by reductive amination reaction. Once a ligand is immobilized, the prepared resin can be used for a variety of affinity purification methods involving either batch or column chromatography. The linkage is stable in elution conditions typically used in most affinity chromatography methods, and allow repeated use of resin after proper wasings and regeneration.

Technical:

• Resin – aldehyde-activated crosslinked 4% beaded agarose, Separopore 4B
• Ideal for coupling antibodies and other proteins – immobilizes proteins via primary amines (-NH2)
• Coupling conditions – very efficient (>88%) coupling over a wide range of pH (4.5-10) and buffer conditions (PBS or bicarbonate buffer with or without organic solvent, DO NOT USE Amine Buffers.
• Stable immobilization – Coupling esults in stable, leak-resistant secondary amine bond between resin and the ligand.
• Versatile and reusable – prepared affinity resin is adaptable to column and batch affinity techniques and the resin is reusable for typical applications based on protein binding interactions
• Convenient sizes and kit –kit containing complete sets of buffers, reagents and versatile spin/drip columns is available (SKU# -------, or select bulk resin.

Product Information:

The Amino coupling reaction involves formation of Schiff base bonds between aldehydes (on the support) and amines (on the ligand) and their subsequent stabilization by incubation with a mild reductant (sodium cyanoborohydride). The entire coupling reaction, called reductive amination occurs in 4 to 6 hours in simple non-amine buffers such as PBS. Coupling efficiency with antibodies and typical proteins is generally greater than 85%, resulting in 1 to 20mg of immobilized protein per milliliter of agarose resin.

Applications: Aldehyde Separopore® 4B has a robust reductive amino alkylation coupling chemistry to immobilize proteins and other ligands through primary amines (–NH₂) to the resin. It can be used for a variety of affinity purification methods involving batch or column chromatography. The resin and linkage are stable in binding and elution conditions.

Note: Separopore® is a cost-effective equivalent to Sepharose® in all of its physical properties and binding characteristics.

Usage: bioWORLD's products are supplied for LABORATORY RESEARCH USE ONLY. The product may not be used as a drug, agricultural or pesticidal product , food additive or as a household chemical.

References

Purification to apparent homogeneity by immunoaffinity chromatography and partial characterization of the GM3 ganglioside-forming enzyme, CMP-sialic acid: lactosylceramide alpha 2,3-sialyltransferase (SAT-1), from rat liver Golgi. J Biol Chem. (1991) 266: 4448-57.
Production of immobilized penicillin acylase using aqueous polymer systems for enzyme purification and in situ immobilization. Enzyme Microb Technol. (2001) 28: 218-224.
Immobilization-stabilization of alpha-chymotrypsin by covalent attachment to aldehyde-agarose gels. Biotechnol Bioeng. (1991) 38: 1144-52.
Stabilization of trypsin by multiple-point attachment to aldehyde-agarose gels. Ann N Y Acad Sci. (1987) 501: 67-72.