Question

This is a question about the SDS - PAGE method, one of the protein analysis methods.

1. I would like to know the functions of resolving gel and stacking gel in SDS - PAGE analysis.

2. I would like to know the various reagents used in manufacturing resolving gel and stacking gel in SDS - PAGE analysis and their role.

We would appreciate your detailed explanation of each question.

Answer 1

Function of resolving gel

This gel used for or capable of allows to separate the proteins and this separation of protein on the basis of their molecular weight.

Function of stacking gel

In the process of separation of protein is needed to concentrate all molecules in the one band therefore they migrating in running gel all the same time. Means by this gel all molecules runs in one band.

Manufacturing of resolving gel

- 10% acrylamide

- 425mM Tris HCL ( pH 8.8)

Manufacturing of stacking gel

- 4% acrylamide

- 125 mM Tris HCL ( pH 6.8)