Isoelectric Focusing is a high-resolution technique for separating
amphoteric molecules. It usually uses acrylamide or agarose gel as support
medium. Amphoteric moleculues such as proteins, enzymes and peptides are
separated on a stable and continuous pH gradient. Isoelectric Focusing
has two stages for separation: At the first stage, carrier ampholytes produce
a pH gradient when they are loaded on the support medium. The current in
the electric field makes the ampholytes to be arranged in a gradient according
to their Isoelectric points. At the second stage, amphoteric molecules
migrate along the pH gradient and finally reach their respective Isoelectric
points.
Followings are Isoelectric Focusing gels run by our laboratory- Laboratory
of Forage Legume Breeding and Genetics, Department
of Agronomy and Soils, Auburn University.
Phosphoglucoisomerase
(GPI) E.C. 5.3.1.9
Esterases
(EST) E.C. 3.1.1.-
Phosphoglucomutase
(PGM) E.C. 2.7.5.1
NAD-dehydrogenase
(Diaphorase, DIA) E.C. 1.6.99.1
Peroxidase
(PRX) E.C. 1.11.1.7
Malic Enzyme
(ME) E.C. 1.1.1.40
Acid phosphatase
(ACP) E.C. 3.2.1.30
6-Phosphogluonate
dehygrogenase (6-PGD) E.C. 1.1.1.44
Superoxide
dismutase (SOD) E.C. 1.15.1.1