Iso Electric focusing (IEF) is a technique that utilises the isoelectric point (pI), for separating molecules.it is focused on a zone of isoelectric point that separates the molecules based on their isoelectric points. The molecules are from the ampholytic source that are added into immobilised pH gradient (IPG) gels. They are acrylamide gel matrix co-polymerised with the pH gradient. This immobilised pH is obtained by continuous change in immobiles ratio. A protein with pH below iso electric point, will be positive and migrates toward cathode through the gradient of increasing pH with changes in protein that ultimately reaches to the it’s corresponding pI. As a result the proteins are changed to sharp stationary bands with each protein positioned at its corresponding pI. After this point there is no net charge and the migration stops. The proteins are Sethu so positioned at their pI ranges that become sharp stationary bands.
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