Notes: Normal No spacing ing 1 Hea Week 7 Post-Lab Questions Name 1) In today\'s

Question

Notes: Normal No spacing ing 1 Hea Week 7 Post-Lab Questions Name 1) In today's lab you loaded 3samples of protein onto the gel, the plasma sample, the flow through fraction and your purified antibody. For each of these samples predict whether you expect it to contain the antibody proteins, and whether you expect it to contain other cellular proteins a. The plasma sample b. The flow through fraction c. Purified antibody 2). In your own words explain why proteins move through the SDS-PAGE gel when they are exposed to an electrical charge. 3) Which protein would you expect to find closer to the wells where the sample was loaded: a protein with a molecular weight of 10,000 Da or a 75,000 Da protein? Explain your answer.

Explanation / Answer

1. a. The plasma sample will contain antibodies depending upon the blood type. The blood of B group will have B antigen on the surface of its red blood cells and A type antibodies in the plasma.

b. Flow through column contains the material that does not bind and hence it is miscellaneous. Hence this may contain many other proteins apart from antibody proteins.

c. Purified antibody will obviously have antibody proteins.

2. Since the SDS also coats the protein with a uniform negative charge these proteins move through the SDS-PAGE gel when exposed to an electrical field they move towards the anode.

3. The protein of higher molecular weight will be closer to the well as it has less mobility as SDS coats the protein with charge uniformly and the proteins are separated solely on the basis of size. Hence 75, 000 Da protein with a higher molecular weight will have less mobility and hence be closer to the well.

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