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Polyacrylamide Gel Electrophoresis- PAGE (Amrita University)

PAGE (Polyacrylamide Gel Electrophoresis), is the most widely used analytical method to resolve separate components of a protein mixture based on size. For this protein molecules of different shapes and sizes, need to be denatured. This is done with the aid of SDS, so that the proteins no longer have any secondary, tertiary or quaternary structure. The proteins covered by SDS are negatively charged. When loaded onto a gel matrix and placed in an electric field, the proteins will migrate towards the anode (positively charged electrode). They are then separated by a molecular sieving effect based on size. After visualization by a protein-specific staining technique, the size of a protein can be estimated by comparison of its migration distance with that of a known molecular weight marker.
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