The Carbohydrate Coupling Resin is designed for the simple and efficient coupling of glycoproteins to a solid agarose support through oxidized sugar groups. The coupling is through the hydrazide group that is immobilized to agarose beads. The resin is ideal for immobilized polyclonal antibodies as these are abundant in carbohydrates in their Fc domain. The location in the Fc domain ensures the antibody’s binding site is orientated away from the resin for optimal binding and reduced steric hindrance.
Glycoprotein sugar components are first oxidized with sodium meta-periodate to convert the cis-glycol groups to reactive aldehydes. These aldehydes react spontaneously with the hydrazide group on the Carbohydrate Coupling Resin, forming stable hydrazone bonds (see figure). The long spacer arm (23Å) reduces steric hindrance and ensures greater binding of proteins and antibodies during affinity purification. The generated affinity resin can be reused at least 10 times with no significant loss of activity.
The Carbohydrate Immobilization kit is supplied with carbohydrate coupling resin columns for the preparation of 2ml affinity columns, a coupling catalyst, SpinOUT™ GT-600 Desalting Columns for removing oxidizing reagents, the oxidizing reagent (Sodium meta-periodate) and relevant buffers.
- Hydrazide activated resin for binding reactive aldehydes
- Couple 1-5mg glycoprotein/ml resin
- Long Spacer Arm (23Å)
- Gentle coupling conditions
- Immobilize glycoproteins
- Immobilize polyclonal antibodies through their Fc domain for optimal orientation
- Immobilize any molecule with oxidized sugars