Describe what type of gel should be used to visualize the fo

Describe what type of gel should be used to visualize the following groups of proteins. Make sure to describe how that type of gel works and why you chose it. Then describe how the protein of choice could be purified. Make sure to describe every step of the purification process with your reasoning.

a) Protein A is 30kDa with a pI of 6. Protein B is 60kDa with a pI of 6. Purify protein B.

b) Protein C is 20kDa with a pI of 4. Protein D is 20kda with a pI of 7. Purify protein C.

c) Protein E is 80kda with a pI of 8 and binds glucose. Protein F is 40kDa with a pI of 5 and does not bind glucose. Protein G is 80kDa with a pI of 5 and binds glucose. Purify protein G.

Solution

A)

we will use SDS-PAGE to purify the protein because the pI of both the proteins are same means charge is same on both proteins and size is different so we will separate both proteins on the basis of there size and we will get two different bands in gel.

B)

we will use Isoelectric focusing to purify the protein because the pI of both the proteins is different means charge is different on both proteins and size is same so we will separate both proteins on the basis of there charge so we can purify them by using Isoelectric focusing (IEF)

C)

We need to perform 2D gel electrophoresis because the charge and size both are different in the mixture. the Mixtures of proteins are separated by two properties in two dimensions on 2D gels. first we will separate protein on the basis of thier charge by using IEF then on the basis of size by using SDS-PAGE