Hydrazide-activated SepFast

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Introduction


The base matrix is activated with hydrazide through a long hydrophilic spacer arm. Hydrazide-activated agarose beads permit the coupling of aldehyde or ketone containing ligands through the formation of stable hydrazone linkages. It is a particularly powerful way to immobilize glycoproteins via their carbohydrate chains while leaving their critical active sites intact. It is a method so called “site-directed immobilization”. For example, antibodies can be immobilized to an agarose solid support through a linkage to their glycosylated side chain, while leaving their Fab regions free for antigen binding.

Glycoproteins can be readily oxidized with sodium periodate to generate formyl groups on their carbohydrate chains before the coupling reaction with Hydrazide-activated agarose beads. It is a successful and well-documented technique. The coupling reaction is mild and easy to carry out. No toxic chemicals or special equipment is required.

Hydrazide activated SepFast 4HF is made of highly cross-linked 4% beaded agarose. It shows high mechanical rigidity allowing high flow throughput with reduced back pressure.

Agarose has long been used for chromatographic separations due to its excellent hydrophilic and low non-specific-binding nature. The particle has open pore structure with excellent mass transfer property to large protein molecules.

Hydrazide-activated SepFast resins can be readily employed to make custom affinity chromatography media for both small scale and large scale purification applications.

Coupling manual is supplied with each product.

Characteristics of Hydrazide-activated SepFast 4HF

 
Group to be coupled -HC=O (aldehyde), -C=O (ketone)
Coupling conditions pH 6 - 10, 4oC to 30oC, 2-16 h, organic solvents possible
Matrix SepFast 4HF: highly cross-linked 4% agarose beads
Particle size 50 – 150 μm
Coupling level >2 mg hIgG / ml medium
pH stability 3 -13 (ligand dependent)
Chemical stability Stable to all commonly used aqueous chemicals, provided the ligand to be coupled can withstand
Storage 4oC – 8oC

Ordering information

 
Product Quantity Code no. Price GBP
Hydrazide-activated SepFast 4HF 5 ml 360101-5ML 81.48
  50 ml 360101-50ML POR
  1 Litre 360101-1L POR

How to order

Operation Manual

Operation Manual