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 |