Lab 9: SDS-PAGE Flashcards
1
Q
Can a protein sample be directly loaded into the gel?
A
No, it needs to be prepared first
2
Q
What are the 3 steps required to prepare a protein to be run on a gel?
A
- Boiling the sample to denature the proteins
- Adding SDS that also denatures and adds an even negative charge around the whole protein
- Adding beta-mercaptoethanol to break the disulfide bonds
3
Q
What is the gel matrix made of for PAGE?
A
Two forms of acryamide that polymerize once two cross-linking agents are added
4
Q
What are the two types of gels used for PAGE?
A
Stacking gel and running gel
5
Q
What is the purpose of the stacking gel?
A
It compresses the samples so they all enter the running gel at the same time
6
Q
What is the purpose of the running gel?
A
Separates the proteins based on size
7
Q
When do we use Coomassie staining to visualize the proteins on the gel?
A
When we want to see all proteins, or looking at one highly abundant protein
