SEPHADEX G-100

Description

Sephadex® G-100 is a gel filtration medium used in protein chromatography, affinity chromatography and gel filtration chromatography.
Sephadex G-100 is useful for desalting and buffer exchange of very large biomolecules.

1. Quickly desalts, removes contaminants and transfers to a new buffer in a single step.
2. Classic gel filtration medium.

Sephadex G-10 is a well established gel filtration medium for desalting and buffer exchange of peptides and small biomolecules >700 molecular weight. Smallest exclusion limit in the Sephadex range.

1. Quickly desalts, removes contaminants and transfers to a new buffer in a single step.
2. Suitable for buffer exchange and cleanup of biological samples, for example peptides, small proteins, and oligosaccharides larger than 700 molecular weight
3. Efficient removal of contaminants such as salts, dyes, and radioactive labels
4. Lowest exclusion limit (700 molecular weight) in the Sephadex G-range

Sephadex is a gel filtration medium prepared by crosslinking dextran with epichlorohydrin. Different types of Sephadex differ in their degree of cross-linking and hence in their degree of swelling and their molecular fractionation range. Sephadex G-10 is one of five different G-types ranging from G-10 for small molecules to G-75 for larger molecules. Sephadex G-50 is available in 4 different particle sizes (Coarse, Medium, Fine & Superfine) and Superfine has the smallest bead size for higher efficiency fractionation with shorter diffusion distances. Coarse and Medium are are preferred for large scale group separations where high flow rates and low operating pressures are required.
Sephadex is a registered trademark of GE Healthcare

History

The introduction of Sephadex in 1959 heralded a new era in separation science. The concept of gel permeation chromatography (GPC) developed slowly and the key events leading to the discovery of Sephadex may be traced to Ingelman ?s early studies on the cross-linking of dextran, the subsequent patent by Flodin and Ingelman on the preparation of cross-linked gels and the exploitation of their gel filtration properties by Flodin and Porath.
Perhaps the most critical event was the synthesis of these gels in bead form with considerably improved flow rates, an innovation that led to the immediate acceptance of these gels in separation technology. Several types of Sephadex are currently available, each with a characteristic fractionation range. The most porous gel, Sephadex G-200, will fractionate proteins in the Mw range 4000–800000, whereas the upper limit for Sephadex G-25 is 5000. Although Sephadex gels are widely used for fractionation and purification of biopolymers, desalting constitutes one of the most important applications. In this context, desalting also refers to buffer exchange operations and the removal of low molecular weight impurities other than salts. Sephadex G-25 is now used for desalting or buffer exchange in many large scale operations. Insulin producers use Sephadex G-50 to remove proinsulin and protease impurities in the final stages of purification of porcine or bovine insulins.
The structure of Sephadex G-25 has recently been investigated by Holmberg (285) and the following novel structural features have come to light.

1. Only 4% of the glucose units are cross-linked.
2. Eight different structural elements were identified, including several glyceryl ethers of glucose and cyclic structures containing 1,4-dioxane rings. Two of these fragments are represented below 3 and 4.
3. 80% of the beads consists of dextran.

Uses

Sephadex^?G-100 is a gel filtration medium used in protein chromatography, affinity chromatography and gel filtration chromatography
It has been used:

• in gel filtration column for α-amylase purification
• in gel filtration column for the purification of polysaccharidic fraction extracted from the skin of the ray Raja radula
• to remove excess protamine by size exclusion from cross-linked iron oxide (CLIO)-protamine-linked (B) peptides

Uses

Fractionation Range (MW)
Globular Proteins: 4,000 - 100,000
Dextrans: 1,000 - 100,000

Uses

Sepharose^? 2B has been used for the preparation of virosomes and platelets.

General Description

Sepharose 2B is a well-proven cross-linked agarose gel filtration base matrix and is frequently used for coupling affinity ligands to the matrix. The matrix is not pre-activated and the user performs all steps in coupling.