SDS-PAGE-What is the purpose of bis-acrylamide

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Reference no: EM133200062

SDS-PAGE Lab Questions

1. SDS-PAGE is an acronym referring to 3 specific components (SDS, PA and GE).

Indicate what SDS-PAGE stands for and briefly describe each of these 3 components.

2. What does it mean to "denature" a protein and why is this important for SDS-PAGE?

3. Describe how you denatured the proteins you used for SDS-PAGE.

4. What substance in sample loading buffer is responsible for breaking disulfide bonds?

5. Describe the purpose of stacking gel and resolving gel with two different pHs and percent acrylamide.

6. What are the possible reasons for not being able to polymerize the acrylamide during gel casting?

7. What is the purpose of bis-acrylamide?

8. What are the purpose of APS and TEMED during polymerization?

9. What is the basis for protein staining using Coomassie blue?

10. Describe briefly the basis and method of silver staining of proteins.

11. What is the purpose of loading "protein molecular weight standards" on the gel?

12. Why did you measure the distance each protein traveled into the gel from the top of the resolving gel and not the bottom of the well?

13. Compare the advantage and disadvantage of Coomassie blue stain and silver stain of protein gels.

Reference no: EM133200062

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