Gel Filtration Chromatography
Electrophoresis is a technique that enables the separation and analysis of charged molecules in an electric field. Gel electrophoresis is a ubiquitous technology used for the separation and purification of biomacromolecules and their fragments. The process of gel electrophoresis works because negatively charged molecules move away from the negative pole of the electric current and smaller molecules will move faster than larger molecules. Thus, size separation is achieved within the pool of molecules running through the gel. The gel works in a similar manner to a sieve separating particles by size. Electrophoresis works by using the particles' inherent charge to move them and pass them through the sieve.
Principle
The fundamental principle of gel filtration chromatography is that the components of a mixture can be separated based on size differences between molecules. Gel filtration chromatography employs spherical gel beads with definite porosity as the packing material in the chromatography column. Depending on the elution limit, some molecules are eluted earlier or later when the components of a liquid mixture pass through the chromatography column. Molecules with high molecular weight than the elution limit will elute earlier. In contrast, molecules with low molecular weight or size than the elution limit will elute later. In this way, the particles are separated by gel filtration chromatography.
Steps
The steps of gel filtration chromatography are as follows.
Applications
One of the major advantages of gel filtration chromatography is that separation can be performed under conditions that are specifically designed to maintain the stability and activity of the molecules without compromising resolution. Because of its simplicity, dependability, versatility, and ease of scale-up, gel filtration chromatography plays a prominent role in the field of biomolecule separation.
Gel filtration chromatography is an effective method for separating biomolecules like enzymes, polysaccharides, nucleic acids, proteins, etc.
Gel filtration chromatography can also be used to facilitate the refolding of denatured proteins by careful control of changing buffer conditions.
Gel filtration chromatography can examine the molecular weight of the separated particles.
If you encounter any problems during the separation of biomolecules using gel filtration chromatography, please feel free to contact us. We'd be happy to help you.
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