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Why does separation not occurs in stacking gel in gel electrophoresis and why does separation done in separating gel?

Answer Posted / Asheesh Chaudhry

In stacking gel, proteins are organized into a narrow band due to electrostatic interactions. This process is called stacking because the proteins are 'stacked' on top of each other like coins in a stack. However, during this process, no actual separation occurs. The separation of proteins happens in the separating gel, where the polyacrylamide matrix has larger pores that allow for the separation of protein molecules based on their size.

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