What is the difference between Native and Denaturing PAGE ? What reagents would

Question

What is the difference between Native and Denaturing PAGE ? What reagents would be included in one but not the other and their purpose? Give a scenario where you would use Native and Denaturing PAGE as it relates to protein analysis
What is the difference between Native and Denaturing PAGE ? What reagents would be included in one but not the other and their purpose? Give a scenario where you would use Native and Denaturing PAGE as it relates to protein analysis
What is the difference between Native and Denaturing PAGE ? What reagents would be included in one but not the other and their purpose? Give a scenario where you would use Native and Denaturing PAGE as it relates to protein analysis

Explanation / Answer

Native PAGE: It is used to separate proteins based on their size, shape, and charge.

Denaturing PAGE: It is used to separate proteins solely based on their size.

In denaturing PAGE, SDS and beta-mercaptoethanol are added. SDS imparts a uniform negative charge on the polypeptide so that the overall charge on a polypeptide is proportional to its size. It also denatures and partially linearizes the protein. Beta-mercaptoethanol is used to reduce disulfide bonds (Intra/intermolecular).

If we want to determine the subunit composition of a protein and its exact molecular size, SDS PAGE is used.

Example: A heterotetrameric protein = A2B2

Subunits are held together by disulfide bonds

A = 40 kDa

B = 60 kDa

In Native PAGE, the protein runs approximately at 200 kDa. It shows a single band.

In SDS PAGE, the protein shows two bands (40 kDa and 60 kDa).

Related Questions

Navigate

• Browse (All)
• Browse W
• Subjects

---

---

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.