What information is provided by non-denaturing PAGE that cannot be provided by denaturing PAGE? (polyacrylamide gel...

What information is provided by non-denaturing PAGE that cannot be provided by denaturing PAGE? (polyacrylamide gel electrophoresis)

Expert Solution

PAGE i.e. polyacrylamide gel electrophoresis is a technique used for the separation of proteins, DNA Molecules based on their size, charge or mass. It is of two types Denaturing PAGE and Non-denaturing PAGE.

Denaturing PAGE is also known as Native PAGE in which the structure of the biomolecule say protein is remains intact. No denaturing Reagent and no reducing buffer is added in the gel. So, the structure of protein remains intact and the migration of protein subunits will be based on its size and charge both. The information can be used for the determination of quaternary structure of protein.

Non-denaturing PAGE is accomplished with reducing buffer (DTT) and Denaturing agent (SDS). SDS will impart negative charge to overall structure of Protein and breaks weak interaction between protein subunits. Reducing agent can break disulfide bonds. These traits will break the Native protein into its respective units. Here, the proteins are separated based on polypeptide chain length i.e. molecular weight only. This can be used for determining the protein sequences, for the molecular weight confirmation.

So, non-denaturing PAGE can provide information about charge of protein but denaturing PAGE can't.

gladiator answered 2 years ago

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