Electrofocusing, or isoelectric focusing, is a technique for separating different molecules by their electric charge differences (if they have any charge). It is most commonly used on proteins.
It is a type of zone electrophoresis that takes advantage of the fact that a molecule's charge changes as the pH of its surroundings changes.
Molecules are distributed over a medium that has a pH gradient (usually created by aliphaticampholytes). An electric current is passed through the medium, creating a "positive" and "negative" end. Negatively charged particles migrate through the pH gradient toward the "positive" end while positively charged particles move toward the "negative" end. As a particle moves into a pH that neutralizes its charge, it will stop following the current. Particles of the same initial charge will deposit (or focus) around the same place on the pH gradient.
See also
Alpha 1-antitrypsin where electrofocusing is used for diagnosis of the enzyme phenotype.
Electrofocusing, or isoelectric focusing, is a technique for separating different molecules by their electric charge differences (if they have any charge).
It is a type of zone electrophoresis that takes advantage of the fact that a molecule's charge changes as the pH of its surroundings changes.
Alpha 1-antitrypsin where electrofocusing is used for diagnosis of the enzyme phenotype.
Electrofocusing is a very high resolution technique which separates biomolecules on the basis of their intrinsic charge.
Used chiefly for peptides and proteins, electrofocusing is fast (one half to one third of the time required by traditional electrophoretic separations) and it has a high sample capacity capable of separating nearly 100 samples on one gel.
Systems have been designed to employ pH gradients which are immobilised into the gel matrix during gel polymerization and thereby eliminating gradient drift and providing resolution of biomolecules differing as little as 0.001pH units in their isoelectric point.
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