1. What is the purpose of boiling your protein samples before loading them into
ID: 252640 • Letter: 1
Question
1. What is the purpose of boiling your protein samples before loading them into the polyacrylamide gel?
2. At the end of electrophoresis and Coomassie Blue Standing, would you expect a protein mixture to appear as a single protein band or as multiple protein bands in the polyacrylamide gel? Explain your reasoning.
3. GFP and BFP both consist of 238 amino acids, but differ in the identifies of two amino acids. Assume that both of these proteins have been denatured by boiling and denaturing solution, and are negatively charged from treatment with SDS.
Would you expect GFP and BFP to migrate the same distance during electrophoresis? Explain your reasoning.
Explanation / Answer
ANS-1 The proteins are boiled before loading into the polyacrylamide gel because it help in complete denaturation of the proteins and it also helpful in physical loading of the proteins in acrylamide gel. The DNA content of the sample can also be reduced by boiling and it makes the sample less gummy.
ans-2 at the end of electrophoresis the protein will be seen as single protein band due to seperation of individual protein from one another. the smaller protein band will be at the largest distance from well while the longest protein will be nearest to well.
ans-3 the diffrence of two amino acid will create some diffrence in net charge of the protein so it will show some impact on the running of both the proteins, if the net chrge is diffrent then the proteins will not migrate to same distance.
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