Gel Filtration

Gel filtration is a simple and reliable chromatographic method for separating molecules according to size. Its versatility makes it generally applicable to the purification of all classes of biological substances, including macromolecules not readily fractionated by other techniques. Gel filtration separations fall into two main categories, group separation and fractionation. In group separation, molecules of widely different molecular sizes are separated. A matrix is chosen such that the larger molecules (e.g. proteins) are eluted in the void volume of the column, whereas small molecules are retained in the total volume. Common examples of group separations are buffer exchange and desalting. Fractionation is used for separation of macromolecules of different sizes. In these applications, the molecular sizes of all the components must fall within the fractionation range of the gel. Typical applications of fractionation using gel filtration are purification and molecular weight determination of proteins, peptides, and nucleic acids.

In recent years, GE Healthcare has introduced new media that produce exceptional resolution at higher flow rates and are more resistant to chemicals, pH, and pressure.

Desalting

Desalting at laboratory scale is a well-proven, simple, and fast method that will rapidly remove low molecular weight contaminants, and at the same time, transfer the sample into the desired buffer in a single step.

GE Healthcare offers a range of prepacked chromatography columns and 96-well filter plates that can be used manually, together with a chromatography system, or in high-throughput applications. These products contain Sephadex™ G-25, a gel filtration medium that allows effective removal of low molecular weight substances from antibodies with a molecular weight > 5000.

Desalting provides several advantages over dialysis, which is generally a slow technique requiring large volumes of buffer and carries the risk of losing material during handling.

Gel Filtration Products

Selection guide: Gel Filtration columns and media pdf

Superdex™ - Prepacked Tricorn™ and PC columns

13 µm particles of Superdex is the first choice for high resolution, short run times and high recovery. Superdex 13 µm chromatography mediumn is available in three different fractionation ranges- Superdex 200, Superdex 75 and Superdex Peptide. The media are available in Tricorn high performance columns and in Precision Columns (PC) for for high-resolution micropurification.

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Superdex™ - HiLoad™ prepacked columns and lab packs

Superdex prep grade is a 34 µm particle intended for larger scale applications. Three different selectivities are available, Superdex 200 prep grade, Superdex 75 prep grade and Superdex 30 prep grade. The different media are available in lab packs as well as in prepacked HiLoad columns.

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Superose™ - Prepacked Tricorn™ and PC columns

The small particle size of Superose gel filtration media allows high- resolution separations across exceptional broad molecular- weight ranges. Superose 6 and Superose 12 are available in Tricorn high performance columns and in Precision Columns (PC) for for high-resolution micropurification.

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Superose™ prep grade - Lab packs

Superose 12 prep grade and Superose 6 prep grade have an average particle size of 30 µm. Both media are available in lab-packs and are useful for preparative purification of higher molecular weight proteins.

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Sephacryl™ - HiPrep™ prepacked columns and lab packs

Sephacryl High Resolution (HR) gel filtration media are suitable for fast, high recovery separations at laboratory scale. Sephacryl HR media provide a useful alternative to Superdex prep grade for application that require a slightly broader fractionation range. HiPrep Sephacryl HR columns, prepacked with Sephacryl HR media, provide reliable gel filtration separations. Sephacryl HR media are also available in lab-packs.

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Sephadex™ - lab packs

Sephadex is ideal for rapid group separations such as desalting and buffer exchange. Sephadex are available in different grades and are supplied lyophilized in different pack sizes. Sephadex G-25 and G-10 are also available in pre-packed columns. Please see Desalting columns below.

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Desalting Products

Selection guide Desalting pdf

Operation

Syringe, Pump, Chromatography system

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Pump, chromatography system

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Centrifuge, Vacuum, Gravity flow

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Related products

Gel Filtration Calibration Kits

Two gel filtration calibration kits are available for protein molecular weight determination by gel filtration. The Low Molecular Weight Kit (LMW) contains 5 proteins with molecular weights in the range 6 500 to 75 000 and Blue Dextran 2000. The High Molecular Weight Kit (HMW) contains 5 proteins with molecular weights in the range 44 000 to 669 000 and Blue Dextran.

Selection guide Calibration Kits

Media	Kit(s)
Superose™ 12, Superose™ 6	LMW and HMW
Superdex™ 75, Superdex™ 75 prep grade	LMW
Superdex™ 200, Superdex™ 200 prep grade	LMW and HMW
Sephadex™ G-75, Sephadex™ G-75 SF, Sephadex™ G-100, Sephadex™ G-100 SF	LMW
Sephacryl™ S-100 HR, Sephacryl™ S-200 HR, Sephacryl™ S-300 HR	LMW and HMW

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Gel Filtration

Selection guide

Gel Filtration columns and media pdf

Handbooks

Gel Filtration, Principles and methods pdf
Protein Purification Handbook pdf