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SDS-PAGE Electrophoresis for Protein Separation – An Overview

Nitu Pathak

Abstract


Polyacrylamide gel electrophoresis (PAGE) is a critical method useful for separating components of a protein mixture depending on their size. The procedure depends on the principle that a charged particle will migrate in an electric field towards a cathode with inverse sign. The general electrophoresis procedures cannot be utilized to determine the molecular weight of organic particles in light of the fact that the versatility of a substance in the gel relies upon both charge and size. To beat this, the organic examples should be dealt with so they obtain uniform charge; at that point the electrophoretic versatility depends basically on estimate. For this diverse protein particles with various shapes and sizes, should be denatured so the proteins lost their auxiliary, tertiary or quaternary structure. The proteins being covered by SDS are contrarily charged and when stacked onto a gel and set in an electric field, it will relocate towards the anode are separated by a molecular sieving effect based on size. After the illustration by a recoloring (protein-particular) scheme, the extent of a protein can be figured by comparing its migration distance with that of known atomic weight marker.

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References


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